Journal: Retrovirology

Article Title: Identification of small molecule modulators of HIV-1 Tat and Rev protein accumulation

doi: 10.1186/s12977-017-0330-0

Figure Lengend Snippet: HIV-1 Tat expression can be rescued with proteasome inhibition by MG132. HeLa HIVrtTA∆ Mls cells were incubated with 791 (30 µM), 833 (2 µM), or 892 (15 µM) for 24 h in the absence (−) or presence of (+) of Dox. Eight hours prior to harvest, MG132 was added as indicated to a subset of the samples. a Representative blot showing effect of the compounds on HIV-1 Gag and Tat expression in the presence or absence of proteasome inhibitor MG132, relative to DMSO treatment. GAPDH serves as loading control. b Summary of band intensities of HIV-1 p24 Gag and p14 and p16 Tat with each treatment relative to that of the DMSO control normalized to the corresponding GAPDH bands (N ≥ 3). Error bars depict standard error of the mean and *, **, and *** indicate P values ≤ 0.05, 0.01, and 0.001, respectively. c , d To assess effect of compound addition on p53 expression, lysates were prepared from cells treated with 791 (30 µM), 833 (2 µM), or 892 (15 µM) and blots were probed for p53 and GAPDH. Shown are ( c ) representative blot of results and d summary of quantitation of p53 expression after normalization using GAPDH

Article Snippet: To determine whether the compounds’ effect on HIV-1 gene expression could be reversed by inhibition of the proteasome, compound treatment assay was performed as previously described with the addition of 10 μM MG132 (Sigma-Aldrich) to compound-treated cells 8 h prior to harvesting.

Techniques: Expressing, Inhibition, Incubation, Quantitation Assay

Journal: bioRxiv

Article Title: PB-scope: Contrastive learning of dynamic processing body formation reveals undefined mechanisms of approved compounds

doi: 10.1101/2025.06.14.659731

Figure Lengend Snippet: MG132 induces a continuous reduction in P-bodies over 8 hours, while thapsigargin causes a temporary decrease in their number from 1 to 3 hours, followed by recovery after 4 hours. These findings are consistent with previous findings – .

Article Snippet: The library of 278 FDA-approved kinase inhibitor compounds and small molecular compounds MG132 and thapsigargin were purchased from TargetMol ( ).

Techniques:

Journal: Cell Death & Disease

Article Title: Stabilization of MCL-1 by E3 ligase TRAF4 confers radioresistance

doi: 10.1038/s41419-022-05500-6

Figure Lengend Snippet: A Immunoblotting for MCL-1 expression in TRAF4 knockout CAL27R and SCC25R cells treated with/without IR (2 Gy) for 72 h. B TRAF4 was transiently transfected into TRAF4 knockout CAL27R cells for 24 h, then treated with IR (2 Gy) and cultured for 72 h. Immunoblotting was used to detect MCL-1 and TRAF4 expression. C – E Ectopic expression TRAF4 compromised IR-induced tumor cell growth inhibition. TRAF4 knockout CAL27R cells were transfected with Flag-TRAF4 for 24 h, then treated with IR (2 Gy) and cultured for 72 h. MTS assay was used to determine the cell viability ( C ), plate colony formation assay to analyze the colony formation ( D ) and soft agar assay to assess the anchorage-independent cell growth ( E ). F CAL27R and SCC25R were transfected with siMCL1 for 24 h, followed by IR (2 Gy) treated and cultured for 72 h. Immunoblotting was performed to detect MCL-1 expression. G – I Silencing of MCL-1 inhibited the growth of tumor cells. CAL27R and SCC25R cells were transfected with siMCL1 for 24 h, followed by IR (2 Gy) treated and cultured for 72 h. MTS assay was performed to determine the cell viability ( G ), plate colony formation assay to analyze the colony formation ( H ) and soft agar assay to assess the anchorage-independent cell growth ( I ). J Immunoblotting for MCL-1 expression in CAL27R cells treated with IR (2 Gy) for 72 h, followed by incubation with MG132 (25 µM) for 6 h. K Immunoblotting for MCL-1 expression in CAL27R cells treated with IR (2 Gy) and incubated with CHX for different time points. L TRAF4 knockout CAL27R cells were treated with MG132 (25 μM) for 6 h, then with IR (2 Gy) for 1 h. Cell extracts were subjected to immunoprecipitation (IP) assay to analyze MCL-1 ubiquitination level. M 293T cells were transfected with HA-Ub, HA-Ub-K48, and HA-Ub-K48R for 48 h, followed by IR (2 Gy) treated for 20 min. IP assay was performed to detect MCL-1 ubiquitination level. L.E long exposure, S.E short exposure. All data are means ± s.e.m. *** p < 0.001, a significant difference between groups as indicated.

Article Snippet: The compounds MG132, cycloheximide (CHX), SB216763 (#S1075), and S63845 were obtained from Selleck Chemicals (Houston, TX).

Techniques: Western Blot, Expressing, Knock-Out, Transfection, Cell Culture, Inhibition, MTS Assay, Colony Assay, Soft Agar Assay, Incubation, Immunoprecipitation, Ubiquitin Proteomics