p38 inhibitor  (TargetMol)

Journal: Cell Death & Disease

Article Title: Repurposing MDM2 inhibitor RG7388 for TP53 -mutant NSCLC: a p53-independent pyroptotic mechanism via ROS/p-p38/NOXA/caspase-3/GSDME axis

doi: 10.1038/s41419-025-07770-2

Figure Lengend Snippet: A , B Detection of ROS by immunofluorescence of HCC827 ( A ) or H23 ( B ) cells stably transfected with non-targeting sgRNA or sgRNA-NOXA. ROS staining (green) and mitochondrial staining (red) are shown. C – E Gene ontology (GO) enrichment analysis using differentially expressed genes showed strong clustering of transcription factor AP-1 complex genes ( JUND, JUNB, JUN and FOS ) which were altered in RG7388 treated cells compared with control cells in HCC827 and PC9 cells. F – K Immunoblot analysis of p-p38, p-JNK, NOXA and β-actin expression in HCC827 or PC9 cells pre-treated with total ROS inhibitor NAC ( F ) or lipid ROS inhibitor Fer-1 ( I ) at the indicated concentration for 1 hour and then treated with 60 μM RG7388 for 6 hours. The protein expression levels of p-p38, p-JNK, and NOXA in HCC827 ( G , J ) or PC9 ( H , K ) were semi-quantitatively assessed using ImageJ software. Quantitative data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. L Immunoblot analysis of p-JNK, NOXA and β-actin expression in HCC827, PC9 or H23 cells pre-treated with 40 μM JNK inhibitor SP600125 for 1 hour and then treated with 60 μM RG7388 for 6 hours. M Immunoblot analysis of p-p38, NOXA and β-actin expression in HCC827 or H1975 cells treated with p38 inhibitor SB203580 (40 μM) for 6 h. N – P Immunoblot analysis of p-p38, NOXA and β-actin expression in HCC827 or PC9 cells pre-treated with 40 μM SB203580 for 1 hour and then treated with 60 μM RG7388 for 6 h. The protein expression levels of p-p38 and NOXA in HCC827 ( O ) or PC9 ( P ) were semi-quantitatively assessed using ImageJ software. Quantitative data are presented as mean ± SD. *** p < 0.001. Q Representative IF images of NOXA intensity (Red) and nucleus stained with DAPI (blue) in HCC827 cells pre-treated with 40 μM SP600125 or SB203580 for 1 hour and then treated with 60 μM RG7388 for 6 h.

Article Snippet: RG7388 (T6254), SB203580 (T1764) and Ferrostatin-1 (T6500) were obtained from Target Mol (Boston, USA).

Techniques: Immunofluorescence, Stable Transfection, Transfection, Staining, Control, Western Blot, Expressing, Concentration Assay, Software

Journal: Cell Death & Disease

Article Title: Repurposing MDM2 inhibitor RG7388 for TP53 -mutant NSCLC: a p53-independent pyroptotic mechanism via ROS/p-p38/NOXA/caspase-3/GSDME axis

doi: 10.1038/s41419-025-07770-2

Figure Lengend Snippet: A Representative phase-contrast images of pyroptotic cell death in HCC827 or H23 cells treated by RG7388 (60 μM), SB203580 (40 μM) or the combination of RG7388 and SB203580 for 6 hours. B – E Immunoblot analysis of cleaved PARP, cleaved caspase-3, GSDME, and cleaved GSDME in the indicated cells treated with RG7388 (60 μM), SB203580 (40 μM) or the combination of RG7388 and SB203580 for 6 hours. The protein expression levels of cleaved PARP ( C ), cleaved caspase-3 ( D ), and cleaved GSDME ( E ) were semi-quantitatively assessed using ImageJ software. Quantitative data are presented as mean ± SD. *** p < 0.001. F – H PC9 and HCC827 cells were treated with RG7388 or the combination of RG7388 and SB203580 for 6 hours. The dead cells were detected by Propidium Iodide (PI) staining. Representative images were shown ( F ). The percentage of PI-positive cells was counted (mean ± SE, n = 5 randomly selected microscope fields) ( G , H ). Student’s t-test was used to analyze the data; *** P < 0.001. I , J Annexin V/PI flow cytometry analysis of H1975 cells under RG7388, SB203580 or combined treatment for 6 h ( I ). The percentage of PI-positive cells was counted (mean ± SE, n = 3) ( J ). *** P < 0.001. K PC9 or H1975 cells in 6-well plates were exposed to DMSO, RG7388, SB203580 or the combination of RG7388 and SB203580 for 6 hours. After 3-4 weeks, the cells were fixed and stained. Representative staining images are shown. L Schematic illustration of ROS/p-p38/NOXA/caspase-3 axis in RG7388 induced TP53 mutant NSCLC cell death.

Article Snippet: RG7388 (T6254), SB203580 (T1764) and Ferrostatin-1 (T6500) were obtained from Target Mol (Boston, USA).

Techniques: Western Blot, Expressing, Software, Staining, Microscopy, Flow Cytometry, Mutagenesis