Journal: Cell Death Discovery

Article Title: MAGL targeted PROTAC degrader simultaneously enhances P53 for synergistic treatment of glioblastoma stem cell

doi: 10.1038/s41420-025-02392-1

Figure Lengend Snippet: A Immunoblotting for MAGL expression in X01 GSCs after 24 h treatment with the indicated concentrations of JZL184 or JN-PROTAC. α-tubulin was used as a loading control. B Immunoblotting for MAGL expression in X01 GSCs treated with 0.5 μM of JN-PROTAC for the indicated durations. C Immunoblotting for MAGL expression in 528 GSCs after 24 h treatment with the indicated concentrations of JZL184 or JN-PROTAC. D , E Immunoblotting for MAGL expression in human breast tumor MDA-MB-231 cells ( D ) and murine melanoma B16F10 cells ( E ) after 24 h treatment with the indicated concentrations of JN-PROTAC. F Immunoblotting for MAGL expression in X01 GSCs after 24 h treatment with 500 nM of JN-PROTAC, 10 μM of TAK-243 or 10 μM of MG-132. TAK-243 and MG-132 are UBA1 inhibitor and proteasome inhibitor, respectively. G Immunoblotting for P53 and MAGL expression in X01 cells after 24 h treatment with the indicated concentrations of JN-PROTAC. H Co-IP assay for the PROTAC-induced ternary complex formation in X01 GSCs treated with JN-PROTAC (10 μM) or DMSO for 48 h. Cell lysates were precipitated with anti-MDM2 antibody. Vinculin was used as a loading control. I Co-IP assay for the interaction of MDM2 and P53 in X01 GSCs treated with JN-PROTAC (10 μM) or DMSO for 48 h. Cell lysates were precipitated with anti-MDM2 antibody. Vinculin was used as a loading control. J The proposed dual functional role of the JN-PROTAC in MAGL degradation and activation of tumor suppressor P53 through MDM2 blocking.

Article Snippet: MG-132 (Cat. No. T2154), and TAK-243(Cat. No. T16974) were purchased from Shanghai Topscience Co., Ltd., and JZL184 (Cat. No. 3836) was received from Tocris Bioscience.

Techniques: Western Blot, Expressing, Control, Co-Immunoprecipitation Assay, Functional Assay, Activation Assay, Blocking Assay

Journal: International Journal of Molecular Sciences

Article Title: Gasdermin D Cleavage and Cytokine Release, Indicative of Pyroptotic Cell Death, Induced by Ophiobolin A in Breast Cancer Cell Lines

doi: 10.3390/ijms27020618

Figure Lengend Snippet: Ophiobolin A activity is dependent on RIPK1 but not RIPK3. ( A ) Viability of cells treated with 500 nM OpA in presence or absence of necrostatin-1 (100 µM). OpA+Nec-1 compared to OpA by Student’s t -test. ( B ) Viability of cells treated with 500 nM OpA in presence or absence of necrostatin-2 (10 µM). The values presented are relative to the viability of vehicle-treated cells and normalized to 100%. Statistical significance of select pairs highlighted following 1-way ANOVA. ( C ) Confocal images of transiently transfected RIPK3-GFP MDA-MB-231 cells and stably transfected RIPK3-GFP HeLa cells treated with doxycycline (20 μg/mL) for 24 h and with 500 nM OpA or 20 ng/mL TNFα+100 nM Smac/LCL161+20 μM zVad or vehicle for 24 h for visualization of the necrosome. Original magnification 20×; scale bar 20 µm. ( D ) Viability of OpA-treated cells in presence or absence of RIPK3 inhibitors (GSK872, GSK840, GSK843). The values presented are relative to the viability of DMSO-treated cells and normalized to 100%. All groups compared via one-way ANOVA with Tukey’s correction for multiple hypothesis testing. ( E ) Immunoblot analysis of total and phospho-RIPK3, total and phospho-MLKL using lysates of OpA- or TSZ-treated cells. β-Actin was used as an internal control. Cells were treated for 24 h. Viability data are presented as the mean ± S.E.M. from at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = not significant.

Article Snippet: Compounds used include zVAD (Cat#S7023, Selleckchem, Houston, TX, USA), Ferrostatin-1 (Cat#SML-0583, MilliporeSigma, St. Louis, MO, USA), Spautin-1 (Cat#56756910MG, MilliporeSigma), Disulfiram (Cat#HY-B024, Tocris Bioscience, Bristol, UK), Necrostatin-1 (Cat# J65341FPL, ThermoScientific, Waltham, MA, USA), Necrostatin-2 racemate (T7504, TargetMol, Boston, MA, USA), GSK872 (Selleckchem), GSK840 (Selleckchem), and GSK843 (Selleckchem).

Techniques: Activity Assay, Transfection, Stable Transfection, Western Blot, Control