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3-Methyladenine (3-MA) Autophagy/PI3K inhibitor

Name: 3-Methyladenine (3-MA)
Brand: Selleck Chemicals
SKU: S2767
Rating: 5 (957 reviews)

제품 번호: S2767

3-Methyladenine (3-MA) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells. It blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also inhibits autophagosome formation. Solutions are unstable and should be fresh-prepared.

3-Methyladenine (3-MA) PI3K inhibitor Chemical Structure

화학 구조

분자량: 149.15

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품질 관리 (Quality Control)

배치: 순도: 99.97%

99.97

함께 자주 사용되는 제품 3-Methyladenine (3-MA)

Necrostatin-1 (Nec-1)

It and Necrostatin-1 inhibit cell death of bone marrow macrophages (BMDM) induced by LPS/zVAD and PolyI: C/zVAD.

Z-VAD-FMK

This compound and Z-VAD-FMK combination confirm vital role of programmed cell death in pristimerin-mediated anti-cancer actions.

Ferrostatin-1 (Fer-1)

This compound and Ferrostatin-1 use abolishes acrylamide (ACR)-induced cell death in chondrocytes.

관련 타겟	Akt mTOR GSK-3 ATM/ATR DNA-PK AMPK PDPK1 PTEN PP2A PDK
기타 PI3K 억제제	GDC-0077 (Inavolisib) SAR405 Quercetin (Sophoretin) LY294002 XL147 analogue Tersolisib (STX-478) Buparlisib (BKM120) 740 Y-P (PDGFR 740Y-P) GO-203 TFA Eganelisib (IPI-549)

세포 배양, 처리 및 작업 농도
(Cell Culture, Treatment & Working Concentration)

세포주	분석 유형	농도	배양 시간	제형	활성 설명	PMID
K562	Function Assay	10mM	1h		decreases the expression of LC3-II and the formation of autophagosomes	21864037
Jurkat	Function Assay	10mM	1h		decreases the expression of LC3-II and the formation of autophagosomes	21864037
HeLa	Cytotoxicity Assay	2mM	24h		inhibites the cytotoxicity of silibinin to HeLa cells.	21875385
PC12/TetOn	Function Assay	0.1/1mM	18h		leads to α-syn(WT) accumulation, toxicity, and oligomer formation	21906659
RMPI8226	Function Assay	5mM	1h		suppresses the level of autophagy under nutrient depletion	21915620
MCF-7	Function Assay	10mM	48h		blocks autophagy induced by bortezomib	21931937
HBx	Apoptosis Assay	10mM	48h	DMSO	increases cell death	22020078
Marc-145	Function Assay	5mM	12/24/36h		reduces the PRRSV titers and the protein expression	22119900
U937	Function Assay	2mM	12h		decreases the autophagy ratio	22155150
BGC-823	Function Assay	5mM	2h		inhibits the rate of autophagic cells	22322152
A549	Function Assay	0.1mM	24h		suppresses SU11274-induced cell death	22466960
pDCs	Function Assay	10mM	0.5h		reduces the induction of IFN-α by ssRNA40	22396599
HeLa	Apoptosis Assay	5mM	24h		induces caspase-dependent cell death	22545128
U251	Apoptosis Assay	5mM	24h		increases S1-induced cell death	22579788
MCF-7	Apoptosis Assay	0.1mM	6h		enhances sirtinol-induced apoptosis	22751989
PC-3	Apoptosis Assay	2mM	2h		increases ORI-induced cell death	22745580
HCT116	Apoptosis Assay	5mM	24h	DMSO	enhances apigenin-induced cell death	24626522
U2OS	Growth Inhibition Assay	10mM	24h		intensifies the growth inhibition induced by Dox	24639013
A2780cp	Apoptosis Assay	2.5mM	1h	ddH2O	enhances cisplatin-induced cell death	24817946
HepG2	Function Assay	5mM	4h		increases cellular levels of HL	24713587
Microglia	Apoptosis Assay	5mM	24h		decreases hypoxia-induced cell death	24818601
MDA-MB 231	Apoptosis Assay	5mM	0.5h		modulates Tocomin® induced apoptosis	24830781
PANC-1	Apoptosis Assay	1mM	48h	DMSO	enhances bortezomib-induced cell viability loss	24842158
MDA-MB-231	Function Assay	2mM	48h		promotes TM-induced cell death	24970676
MDA-MB-231	Function Assay	2mM	24h		inhibits autophagy induced by TM	24970676
MCF-7	Function Assay	2mM	48h		promotes TM-induced cell death	24970676
MCF-7	Function Assay	2mM	24h		inhibits autophagy induced by TM	24970676
HepG2	Apoptosis Assay	3mM	5h		reduces cell apoptosis induced by QDs	22836595
HeLa	Apoptosis Assay	10mM	2h		decreases cell viability co-treatment with PEI	23000135
SK-HEP-1	Apoptosis Assay	10mM	1h		protects against autophagy and induces apoptosis in bufalin-treated cells	22858649
MDA-MB231	Function Assay	5mM	1h		increases resveratrol-mediated caspase activation and cell death	23088850
PaCa44	Apoptosis Assay	2.5mM	1h		reduces genipin-mediated apoptosis	23124112
T-47D	Function Assay	10mM	2h		inhibits autophagy process and increases rapamycin induced apoptosis	23300026
GTL-16	Apoptosis Assay	5mM	24h		reduces cell viability as compared to cells treated with MET inhibitors	23313490
U251MG	Function Assay	3mM	1h		suppresses LC3-II protein expression	23338618
T24	Function Assay	10mM	1h		reduces the cleavage of LC3 after baicalin treatment	23354080
HUVECs	Function Assay	3mM	24h		blocks the protective effect of resveratrol by inhibiting autophagy	23358928
MCF-7	Function Assay	5mM	24h		inhibits starvation-induced autophagy	23395679
Hela	Function Assay	5mM	24h		inhibits starvation-induced autophagy	23395679
OR6	Function Assay	10mM	72h		suppresses HCV replication and formation of autophagosomes	23395875
HT-29	Function Assay	1mM	48/96h		inhibits AMPK induces autophagic cell death	23508272
SH-SY5Y	Cytotoxicity Assay	5mM	24h		increases PCN toxicity	23525265
Saos-2	Apoptosis Assay	1mM	96h		increases cell death induced by PCX	23563171
1321N1	Cytotoxicity Assay	5mM	24h		protects cell against PCN-induced toxicity	23525265
A2780	Apoptosis Assay	5mM	24h		converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells	23592281
OV2008	Apoptosis Assay	5mM	24h		converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells	23592281
PC12	Function Assay	10mM	24h	water	inhibits chymotrypsin-like proteasomal activity.	23603979
SH-SY5Y	Apoptosis Assay	5mM	1h		abolishes celastrol neuroprotective effect	23619395
SH-SY5Y	Function Assay	1mM	24h		inhibits the autophagy induced by TOCP	23743148
HepG2	Function Assay	10mM	24h		inhibits siTIGAR- and HBSS-induced autophagy	23817040
HeLa	Function Assay	10mM	2h		suppresses LC3 II expressison	23864738
HONE-1	Function Assay	5mM	1h		represses 6r-mediated ROS production	23892358
MCF7	Function Assay	5mM	24h		increases CuO induced cell death	23962629
HO8910	Apoptosis Assay	10mM	12h		enhances B19-induced apoptosi	23983610
SMMC-7721	Apoptosis Assay	5mM	24h		attenuates TNF-α protection against serum starvation-mediated apoptosis	24066693
Hep3B	Apoptosis Assay	5mM	24h		attenuates TNF-α protection against serum starvation-mediated apoptosis	24066693
H460	Function Assay	10mM	4h		increases cisplatin-induced cell death	24173208
A549	Function Assay	10mM	4h		inhibits autophagy induced by irradiation	24142735
H1299	Function Assay	10mM	4h		increases cisplatin-induced cell death	24173208
WiDr	Function Assay	10mM	1h		inhibits PCBL-induced LC3 II expression	24190489
LoVo	Apoptosis Assay	5mM	48h		enhances DCA-induced apoptosis.	24201812
HepG2 E47	Function Assay	2.5mM	48h		increases the toxicity of AA, BSO, and CCl4	24273738
RKO	Function Assay	2mM	1h	DMSO	enhances cell death by geldanamycin	24291777
Hep3B	Apoptosis Assay	2mM	12h	DMSO	inhibits AZD8055-induced cell death	24297300
ACHN-5968	Apoptosis Assay	5mM	3h		enhances paclitaxel-mediated apoptosis	24305604
Huh7	Apoptosis Assay	2mM	12h	DMSO	inhibits AZD8055-induced cell death	24297300
UOK257	Apoptosis Assay	5mM	3h		enhances paclitaxel-mediated apoptosis	24305604
ECSCs	Apoptosis Assay	10mM	4h		decreases rapamycin-treated apoptosis	24319109
MCF-7	Function Assay	10mM	24h		inhibits the autophagy induced by chemotherapy drugs	24315578
SGC-7901	Apoptosis Assay	2mM	1h		increases CA-4 induced apoptosis	24321340
SMMC-7721	Apoptosis Assay	2mM	1h		increases CA-4 induced apoptosis	24321340
T24	Apoptosis Assay	5mM	1.5h		potentiates celecoxib-induced apoptosis	24349176
NTUB1	Apoptosis Assay	5mM	1.5h		potentiates celecoxib-induced apoptosis	24349176
MG-63	Apoptosis Assay	10mM	12h		enhances DP-induced apoptosis	24358301
MG-63	Apoptosis Assay	0.5/1mM	32h		enhances salinomycin-induced cell apoptosis	24358342
MG-63	Function Assay	0.5/1mM	48h		induces salinomycin-induced cell viability loss	24358342
U2OS	Function Assay	0.5/1mM	48h		induces salinomycin-induced cell viability loss	24358342
HGC-27	Function Assay	10mM	1h		inhibits the cell viability loss by RAD001 or MK-2206	24416349
HCT116	Apoptosis Assay	5mM	24h		enhances the apoptosis induced by apigenin	24626522
A549	Apoptosis Assay	10mM	48h		accelerates the reduction of cell viability induced by PTX	24626722
Saos-2	Apoptosis Assay	10mM	24h		intensifies the growth inhibition of the U2OS cells induced by Dox	24639013
U2OS	Apoptosis Assay	10mM	24h		intensifies the growth inhibition of the U2OS cells induced by Dox	24639013
HepG2	Function Assay	5mM	4h		increases HL release	24713587
A549	Apoptosis Assay	5mM	48h		decreases the percentage of cell death and expression levels of caspase-3, Beclin-1 and LC3-II	24706303
A2780cp	Apoptosis Assay	2.5mM	1h	ddH2O	enhances cisplatin-induced cell death	24817946
Microglia	Apoptosis Assay	5mM	24h		decreases hypoxia-induced cell death	24818601
HT-29	Apoptosis Assay	1mM	48h	DMSO	enhances bortezomib-induced cell viability loss	24842158
MDR	Apoptosis Assay	10mM	6h		strengthens the power of anticancer drugs	25019701
H157	Function Assay	5mM	2h		suppresses SPC induced accumulation of LC3-II	25285628
A549	Function Assay	5mM	2h		suppresses SPC induced accumulation of LC3-II	25285628
A2780cp	Growth Inhibition Assay	1mM	1h		increases cisplatin-induced cell death	25322694
NBL-W-S	Apoptosis Assay	1mM	6h		increases cell apoptosis induced by GANT-61	25323222
NBL-W-S	Growth Inhibition Assay	1mM	6h		enhances GANT-61 toxicity	25323222
A549	Apoptosis Assay	5mM	2h	DMSO	inhibits BDMC-induced apoptotic cell death	25716561
95D	Apoptosis Assay	5mM	2h	DMSO	inhibits BDMC-induced apoptotic cell death	25716561
A549	Growth Inhibition Assay	3mM	2h	DMSO	reduces growth inhibitory effect of BDMC	25716561
95D	Growth Inhibition Assay	3mM	2h	DMSO	reduces growth inhibitory effect of BDMC	25716561
Nara-H	Growth Inhibition Assay	5mM	48h		enhances temsirolimusmediated suppression of Nara-H cell proliferation	21805033
HUVECs	Function Assay	10mM	0.5h		decreases the AGE-BSAinduced autophagy leve	21468592
HepG2	Apoptosis Assay	2mM	1h		enhances radiation-induced cell death	21453691
U-2 OS	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells	29435139
A673	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
DAOY	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells	29435139
Saos-2	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
BT-37	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
RD	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
SK-N-SH	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
BT-12	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells	29435139
MG 63 (6-TG R)	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells	29435139
NB1643	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
Rh41	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
Rh30	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh30 cells	29435139
SJ-GBM2	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
SK-N-MC	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB-EBc1	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
LAN-5	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
Rh18	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells	29435139
클릭하여 더 많은 세포주 실험 데이터 보기

화학 정보, 보관 및 안정성 (Chemical Information, Storage & Stability)

분자량	149.15	화학식	C₆H₇N₅	보관 (수령일로부터)	3 years -20°C powder
CAS 번호	5142-23-4	SDF 다운로드		원액 보관	용액은 불안정합니다. 신선하게 준비하거나 소량의 사전 포장된 크기를 구매하십시오. 수령 즉시 재포장하십시오.
동의어	NSC 66389			Smiles	CN1C=NC(=N)C2=C1N=CN2

용해도 (Solubility)

In vitro
배치:

DMSO : 10 mg/mL (67.04 mM) 60°C 수조에서 가온; 초음파 처리;
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Ethanol : 10 mg/mL

Water : 4 mg/mL

Ethanol : 12 mg/mL

DMSO : 8 mg/mL (53.63 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Water : Insoluble

DMSO : 21 mg/mL (140.79 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Water : 20 mg/mL

Ethanol : 14 mg/mL

DMSO : 16 mg/mL (107.27 mM) 60°C 수조에서 가온; 초음파 처리;
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Water : 10 mg/mL (超声加热十分钟，60℃)

Ethanol : 8 mg/mL

Ethanol : 6 mg/mL

DMSO : 5 mg/mL (33.52 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Water : 3 mg/mL

몰농도 계산기

질량	농도	부피	분자량
=	×	×

희석 계산기 분자량 계산기

In vivo 배치:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Selleck 연구소에서 검증되었습니다. 이 제형에 대한 조정이 필요한 경우 맞춤 테스트를 위해 당사 영업팀에 문의하십시오.	0.750mg/ml (5.03mM)	Taking the 1 mL working solution as an example, add 50 μL of 15 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Selleck 연구소에서 검증되었습니다. 이 제형에 대한 조정이 필요한 경우 맞춤 테스트를 위해 당사 영업팀에 문의하십시오.	0.500mg/ml (3.35mM)	Taking the 1 mL working solution as an example, add 50 μL of 10 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

생체 내 제형 계산기 (투명한 용액)

1단계: 아래 정보 입력 (권장: 실험 중 손실을 고려하여 추가 동물 포함)

투여량: mg/kg 동물 평균 체중: g 동물당 투여량:μL 동물 수:

2단계: 생체 내 제형 입력 (이것은 계산기일 뿐 제형이 아닙니다. 용해도 섹션에 생체 내 제형이 없는 경우 먼저 당사에 문의하십시오.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

계산 결과:

작업 농도: mg/ml;

DMSO 원액 준비 방법: mg 약물 사전 용해 μL DMSO ( 원액 농도 mg/mL, 농도가 해당 약물 배치의 DMSO 용해도를 초과하는 경우 먼저 당사에 문의하십시오. )

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가μL PEG300, 혼합하고 투명하게 한 다음 추가μL Tween 80, 혼합하고 투명하게 한 다음 추가 μL ddH₂O, 혼합하고 투명하게 합니다.

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가 μL 옥수수 기름, 혼합하고 투명하게 합니다.

참고: 1. 다음 용매를 추가하기 전에 액체가 투명한지 확인하십시오.
2. 용매를 순서대로 추가해야 합니다. 다음 용매를 추가하기 전에 이전 추가에서 얻은 용액이 투명한 용액인지 확인해야 합니다. 와동, 초음파 또는 뜨거운 물 중탕과 같은 물리적 방법을 사용하여 용해를 도울 수 있습니다.

작용 메커니즘 (Mechanism of Action)

Targets/IC₅₀/Ki	Autophagy Vps34 (HeLa cells) 25 μM PI3Kγ (HeLa cells) 60 μM
시험관 내(In vitro)	3-Methyladenine (3-MA)이 Vps34를 예방하는 경미한 선호도는 아마도 Vps34에 특이적인 소수성 고리에서 기인하며, 이는 이 화합물의 3-메틸기를 둘러싸고 있습니다. 이는 정상 상태와 기아 상태 모두에서 암세포 사멸을 유발한다고 보고되었으며, Autophagy를 억제하는 능력과 독립적으로 세포 이동 및 침윤을 억제할 수 있어 Autophagy 억제 이외의 기능을 보유하고 있음을 시사합니다. 이 화합물은 Autophagy 억제와 독립적인 caspase 의존적 세포 사멸을 유도합니다. 5 mM의 3-MA로 처리하면 GFP-LC3 점을 나타내는 포도당 결핍 HeLa 세포의 비율이 23%로 감소합니다. 3-MA로 처리된 세포에서 12시간에서 48시간 사이에 LC3-I 수준은 증가하고 LC3-II 수준은 감소합니다. LC3-I에서 LC3-II로의 전환은 이 화합물에 의해 억제됩니다. 2.5 mM 또는 5 mM의 농도로 1일 동안 HeLa 세포를 처리하면 세포 생존율에 영향을 미치지 않지만, 10 mM 농도로 1일 동안 처리하면 세포 생존율이 25.0% 감소합니다. 2.5, 5 또는 10 mM 농도로 2일 동안 처리하면 생존율이 각각 11.5%, 38.0%, 79.4% 감소합니다. 이 화합물은 시간 및 용량 의존적 방식으로 세포 생존율을 감소시키며, nocodazole에 의해 유도된 전중기 정지 기간을 상당히 단축시킵니다. 3-MA에 의한 Autophagy 억제는 SU11274로 유도된 세포 사멸을 저해합니다. 3-MA로 장기간(최대 9시간) 처리하면 야생형 MEF에서 상당한 LC3 I에서 II로의 전환이 유도됩니다. wortmannin이 아닌 3-MA로 장기간 처리하면 GFP-LC3 점/응집이 현저하게 증가합니다. 3-MA로 유도된 LC3 전환 및 유리 GFP 방출은 ATG7에 의존적입니다. 3-MA 처리는 p62 단백질 수준의 명백한 증가로 이어집니다. 이 화합물은 Atg5−/− MEF뿐만 아니라 DOX 매개 ATG5 결실 세포에서도 p62 수준을 증가시킵니다. 이는 서로 다른 시간적 패턴으로 class I PI3K 및 class III PI3K를 억제합니다. 3-MA로 유도된 LC3 I에서 LC3 II로의 전환은 야생형 세포에 비해 Tsc2−/− 세포에서 현저하게 저해됩니다. 이 화합물은 mTOR complex 1의 항-Autophagy 기능을 방해합니다.
키나아제 분석	단백질 분해 분석
키나아제 분석	HeLa 세포를 0.05 mCi/mL l-[U-¹⁴C]valine으로 24시간 동안 방사성 표지합니다. 표지 기간이 끝나면, 세포를 PBS로 3회 세척합니다. 세포는 10 mM 3-Methyladenine (3-MA) 존재 여부에 따라 완전 배지 또는 EBSS에서 지정된 시간 동안 배양됩니다.
생체 내(In vivo)	3-Methyladenine (3-MA)은 class III PI3K에 대한 효과를 통해 Autophagy를 차단합니다. 이 화합물로 처리해도 지주막하 출혈(SAH) 그룹과 비교하여 출혈 정도는 변하지 않습니다. 전처리는 SAH + vehicle 그룹과 비교했을 때 신경학적 증상을 유의하게 악화시킵니다. 이를 적용하면 Autophagy가 감소합니다. 반대로, SAH + 3-MA 그룹에서는 cleaved caspase-3가 현저하게 상향 조절됩니다. cleaved caspase-3 발현의 상향 조절과 일치하게, 우측 피질의 TUNEL 양성 세포 수는 SAH + vehicle 그룹과 비교하여 SAH + 3-MA 그룹에서 유의하게 증가합니다.
참조	[1] https://pubmed.ncbi.nlm.nih.gov/20574157/ [2] https://pubmed.ncbi.nlm.nih.gov/22545128/ [3] https://pubmed.ncbi.nlm.nih.gov/22466960/ [4] https://pubmed.ncbi.nlm.nih.gov/20123989/ [5] https://pubmed.ncbi.nlm.nih.gov/22521819/

적용 분야 (Applications)

방법	바이오마커	PMID
Western blot	APP / BACE1 / ADAM17 / Presenilin 1 / Presenilin 2 / Nicastrin / APH-1 / Pen-2 / LC3-1 / LC3-2 VEGF caspase-3 / caspase-9 / PARP α-SMA / TGF-β / LC-3BI / LC-3B II / Beclin-1 / NF-κB p65	26316755
Immunofluorescence	LC3 / Hif-α / COX2	29039446
Growth inhibition assay	Cell viability	26934124

자주 묻는 질문 (Frequently Asked Questions)

질문 1:
I'm also wondering whether it can be dissolved in water, or maybe something like culture medium, normal saline solution to form 10mM solution.

답변:
As the reference (http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal. pone.0035665), it was found to inhibit autophagy at concentrations ranging from 1 to 10 mM and was directly dissolved into the culture medium at the indicated concentrations. And we tested the solubility of S2767, and found its solubility in DMEM is 31 mg/mL at about 40°C.

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