3-MA effects on protein degradation depend on the experimental setting. In HeLa cells, 10 mM 3-MA reduced long-lived protein degradation during 4 h in EBSS but increased degradation during 8 h in full medium. Autophagy inhibition therefore should not be assumed across nutrient conditions and exposure times. [1]
In MCF-7 and T-47D breast cancer cells, 3 mM 3-MA pretreatment followed by co-incubation reduced the apoptosis associated with 150 μM 18β-glycyrrhetinic acid. Cleaved PARP and LC3-II also decreased. The findings describe this combination under the tested conditions and do not demonstrate anticancer efficacy of 3-MA alone. [2]
Time-dependent effects on cellular PI3K lipid products
Cell-based study · Wild-type mouse embryonic fibroblasts in full DMEM with 10% fetal bovine serum [1]
3-MA reduced PI3P early, with recovery at 9 h, whereas PI(3,4,5)P3 remained suppressed. Cellular lipid endpoints support differential temporal PI3K pathway effects; they are not purified-enzyme inhibition constants.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5 mM
- Exposure duration
- 1, 3 and 9 h (mass strips); 1, 3, 6 and 9 h (chromatography)
- Method
- Addition to culture medium
- Reported limits
- 3-MA stock solvent and purity not specified in reviewed methods. Fig. 5C caption prints wortmannin 50 mM, versus 50 nM in 5A; comparator concentration not normalized or adopted.
Methods and source locations
Compound reported in the paper: 3-Methyladenine; Compound label in the paper: 3-MA
Evidence applies to the named compound in the reported model and conditions; it does not validate an Adooq lot or establish clinical efficacy.
Fig. 5A–C; Results: 3-MA Inhibits Class I and Class III PI3K in Different Temporal Patterns; Methods: lipid quantification, cell culture, reagents
Cellular PI3P abundance: PI3P mass-strip assay with untreated full-medium control, wortmannin comparator and EBSS condition — Fig. 5A, image and caption; corresponding Results
Cellular PI(3,4,5)P3 abundance: PI(3,4,5)P3 mass strips and chromatography; chromatography data mean ± S.D., three independent experiments; Student t test, ** p < 0.01 as annotated — Fig. 5B–C; corresponding Results; Statistical Analysis
Nutrient- and duration-dependent long-lived protein degradation
Cell-based study · HeLa cells in full DMEM with 10% fetal bovine serum or nutrient-deprived EBSS [1]
At 10 mM, 3-MA reduced long-lived protein degradation in EBSS at 4 h but increased degradation in full medium at 8 h. This functional endpoint limits a condition-independent autophagy-inhibitor description.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 mM
- Exposure duration
- 4 h or 8 h, as specified for each condition
- Method
- Addition to culture medium
- Reported limits
- Stock solvent not specified. Different time/medium comparisons do not isolate nutrient status from treatment duration.
Methods and source locations
Compound reported in the paper: 3-Methyladenine; Compound label in the paper: 3-MA
Evidence applies to the named compound in the reported model and conditions; it does not validate an Adooq lot or establish clinical efficacy.
Fig. 2F; Results: 3-MA Increases Autophagic Flux; Methods: Protein Degradation Assay and Cell Culture
Long-lived protein degradation: Radiolabeled valine protein degradation assay; matched untreated controls. Fig. 2F: means ± S.D., three independent experiments; Student t test, ** p < 0.01 for these comparisons — Fig. 2F image/caption and Results: long lived protein degradation paragraph
3-MA attenuates 18β-GA-associated apoptosis
Cell-based study · Human MCF-7 and T-47D breast cancer cells [2]
3-MA decreased apoptosis caused by 18β-glycyrrhetinic acid (18β-GA) in both cell lines. Lower cleaved PARP and LC3-II accompanied the combination. This is a combination effect and does not establish that 3-MA alone is an anticancer treatment or a specific autophagy-only causal test.
Literature-Reported Treatment Conditions
- Treatment concentration
- 3-MA 3 mM (figure labels); 18β-GA 150 μM
- Exposure duration
- 3-MA pretreatment 2 h; subsequent co-incubation 48 h for apoptosis, 24 h for immunoblot
- Method
- Addition to culture medium
- Co-treatment
- 18β-GA 150 μM; rapamycin comparator 30 μM shown in the figure
- Reported limits
- 3-MA-specific vehicle, purity, catalog number and lot not specified. Fig. 5 caption does not specify its experiment count; n from other figures is not transferred.
Methods and source locations
Compound reported in the paper: 3-Methyladenine; Compound label in the paper: 3-MA
Evidence applies to the named compound in the reported model and conditions; it does not validate an Adooq lot or establish clinical efficacy.
Fig. 5A–B, image/caption; Results: autophagy induction and 18β-GA apoptosis; Methods: reagents, apoptosis analysis, Western blotting
Annexin V-FITC/PI apoptosis: Flow cytometry, 48 h; control, 3-MA alone, 18β-GA alone and combination shown; Fig. 5 # p < 0.05 versus 18β-GA, one-way ANOVA/Bonferroni — Fig. 5A–B upper plots and caption; apoptosis methods
Cleaved PARP and LC3-II: Western blot after 24 h; GAPDH reference; directional interpretation from bands, histograms and corresponding Results — Fig. 5A–B immunoblots/histograms and corresponding Results
