Journal: Biochemistry and Biophysics Reports

Article Title: Bifendate inhibits cell PARthanatos by activating the MEK/ERK pathway

doi: 10.1016/j.bbrep.2026.102454

Figure Lengend Snippet: DDB sustains cellular phospho-ERK levels. (A–B) HeLa cells were pre-treated with DDB for 24 h, exposed to MNNG (60 μM, 15 min) and cultured for a further 8 h. ERK1 and phospho-ERK1/2 expression were analysed by immunoblot (A) and densitometry (B). (C) Cells pre-treated with DDB for 24 h were challenged with MNNG (60 μM, 15 min) and harvested 4 h later; MEK1/2 and phospho-MEK1 levels were analysed by immunoblot. (D) The thermal stability of MEK and ERK after 24 h pretreatment with DDB (25 μM) or DMSO was detected by Western blot. (E) The grey values of each protein band were analysed quantitatively. (n = 3). The values are expressed as the means ± SDs. Statistical analysis was performed using T-test. *p < 0.05. (F) The thermal stability of DUSP1 after 24 h pretreatment with DDB (25 μM) or DMSO was detected by Western blot. (G) The grey values of protein band were analysed quantitatively. (n = 3). The values are expressed as the means ± SDs. Statistical analysis was performed using T-test. (H) Cells were pre-treated or not with DDB for 24 h, exposed to MNNG (60 μM, 15 min) and cultured for 4 h then phospho-Bad expression was analysed. (I) Mitochondrial membrane potential was assessed by JC-1 staining after the indicated treatments. Scale bar: 50 μm. The values are expressed as the means ± SDs (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's test. **p < 0.01.

Article Snippet: After sealing with 5 % skimmed milk, the membrane was incubated successively with the primary antibody (PAR/pADPr, R&D, #4335-MC-100; PARP1, ABclonal, #A0010; AIF, Santa Cruz Biotechnology, #sc-13116; TOMM20, Beyotime, #AF1717; MIF, Beyotime, #AF7467; β-Tubulin, Beyotime, #AF2839; ERK1, Beyotime, #AF1315; Phospho-Erk1 (Thr202/Tyr204)/Erk2 (Thr185/Tyr187), Beyotime, #AF1891); MEK1/2, Beyotime, #AF1057; Phospho-MEK1 (Ser218/222), Beyotime, #AF1786; DUSP1, MCE, #HY- P80645 ; GAPDH, ABclonal, # AC002) and the secondary antibody (ABclonal, #AS014; ABclonal, #AS003).

Techniques: Cell Culture, Expressing, Western Blot, Membrane, Staining

Journal: Cell Death & Disease

Article Title: Protein deglycase DJ-1 deficiency aggravates acute viral myocarditis by promoting apoptosis via reducing Dusp1 expression

doi: 10.1038/s41419-025-08185-9

Figure Lengend Snippet: A RNA transcriptome sequencing of the heart from DJ-1 −/− and WT mice. The fold change of all genes with a significant difference was presented in the volcano plot. B KEGG analysis of downregulated genes in DJ-1 −/− mice compared with the WT type. C H9C2 cells were transfected with siRNA targeting DJ-1 (si-DJ-1) or a control scrambled sequence (si-NC). The mRNA levels of DJ-1 and Dusp1 were determined by RT-qPCR at the indicated time. D Quantitative RT-qPCR analysis of Dusp1 mRNA level in the heart of DJ-1 deletion mice and wild-type mice. E – H Depressed protein level of Dusp1 and increased activation of the P38MAPK pathway in the DJ-1 deficiency groups compared with the control (the data are represented as the mean ± SEM. One-way ANOVA by post-test (Tukey) analysis was used. n = 3. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: The following antibodies were used: DJ-1 (Abcam, ab76008), VP1 (GeneTex, GTX132346), Pro-caspase 3 (Cell Signaling Technology, #9662), Cleaved caspase 3(Cell Signaling Technology, #9664), Dusp1 (Santa, sc-373841), P38MAPK (Cell Signaling Technology, #8690), pP38MAPK (Cell Signaling Technology, #4511), Gapdh polyclonal antibody (Proteintech, No. 10494-1-AP).

Techniques: Sequencing, Transfection, Control, Quantitative RT-PCR, Activation Assay

Journal: Cell Death & Disease

Article Title: Protein deglycase DJ-1 deficiency aggravates acute viral myocarditis by promoting apoptosis via reducing Dusp1 expression

doi: 10.1038/s41419-025-08185-9

Figure Lengend Snippet: A , B Western blot analyses revealed that DJ-1 deficiency downregulated Dusp1 upon CVB3 infection in the heart of mice. The expression levels of DJ-1 and Dusp1 in hearts from DJ-1 −/− and WT mice were assessed by western blotting. C , D Flow cytometry was used to detect the apoptosis rate. E , F The expression levels of DJ-1, Dusp1, P38 MAPK, pP38 MAPK, Pro-caspase-3, and Cleaved caspase-3 were assessed by western blotting. Lv-Dusp1, H9C2 cells transfected with lentivirus encoding Dusp1. Lv-NC, H9C2 cells transfected with the lentivirus vector. si-DJ-1, H9C2 cells transfected with siRNA targeting DJ-1. si-NC, H9C2 cells transfected with a control scrambled sequence. The data are represented as the mean ± SEM. n = 3. One-way ANOVA by post-test (Tukey) analysis was used. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: The following antibodies were used: DJ-1 (Abcam, ab76008), VP1 (GeneTex, GTX132346), Pro-caspase 3 (Cell Signaling Technology, #9662), Cleaved caspase 3(Cell Signaling Technology, #9664), Dusp1 (Santa, sc-373841), P38MAPK (Cell Signaling Technology, #8690), pP38MAPK (Cell Signaling Technology, #4511), Gapdh polyclonal antibody (Proteintech, No. 10494-1-AP).

Techniques: Western Blot, Infection, Expressing, Flow Cytometry, Transfection, Plasmid Preparation, Control, Sequencing