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: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Dusp1 mRNA levels on P78. B , DUSP1 protein levels and activity on P78. C , DUSP1 activity on P78. D , DUSP1 staining in CaMKII-positive neurons. Scale bar, 400 μm/100 μm. E , Levels of MAPK-related signal proteins on P78, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ASE-C, subthreshold-dosed administration of cocaine in adolescent saline-exposed mice; ACE-C, subthreshold-dosed administration of cocaine in adolescent cocaine-exposed mice; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ASE-C mice.

Article Snippet: The transferred membranes were blocked with 5% non-fat dry milk and 0.1% Tween 20 in 10 mM Tris–HCl (TBST buffer, pH 7.5) for 1.5 h at room temperature, then subsequently incubated with the following primary antibodies: c-Fos (1:1500, Rabbit, RRID: AB_2247211, Cell Signalling Technology, USA), DUSP1 (1:1000, Rabbit, BM4856, Boster Biotechnology, China), ERK1/2 (1:1000, Rabbit, BM4326, Boster Biotechnology, China), p-ERK1/2 (1:1000, Rabbit, BM4156, Boster Biotechnology, China), CREB (1:1000, Rabbit, RRID:AB_331277, Cell Signaling Technology, USA), p-CREB (1:1000, Rabbit, RRID:AB_2561044, Cell Signaling Technology, USA), BDNF (1:1000, Mouse, RRID:AB_2881675, Proteintech, USA).

Techniques: Activity Assay, Staining, Saline

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , DUSP1 protein levels on P74. B , DUSP1 activity on P74. ASE, adolescent salineexposed mice; ACE, adolescent cocaine-exposed mice; N.S., p > 0.05 vs ASE mice.

Article Snippet: The transferred membranes were blocked with 5% non-fat dry milk and 0.1% Tween 20 in 10 mM Tris–HCl (TBST buffer, pH 7.5) for 1.5 h at room temperature, then subsequently incubated with the following primary antibodies: c-Fos (1:1500, Rabbit, RRID: AB_2247211, Cell Signalling Technology, USA), DUSP1 (1:1000, Rabbit, BM4856, Boster Biotechnology, China), ERK1/2 (1:1000, Rabbit, BM4326, Boster Biotechnology, China), p-ERK1/2 (1:1000, Rabbit, BM4156, Boster Biotechnology, China), CREB (1:1000, Rabbit, RRID:AB_331277, Cell Signaling Technology, USA), p-CREB (1:1000, Rabbit, RRID:AB_2561044, Cell Signaling Technology, USA), BDNF (1:1000, Mouse, RRID:AB_2881675, Proteintech, USA).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Experimental design and timeline. B , Schematic diagram of viral injection. Scale bar, 400 μm. C , Percentage of viral transfected neurons in CaMKII-positive neurons of mPFC. Scale bar, 200 μm/100 μm. D , Dusp1 mRNA levels on P78. E , DUSP1 protein levels on P78. F , DUSP1 activity on P78. G , CPP score. H , ΔCPP score. I , Heatmap of spent duration by mice in CPP apparatus. J , C-Fos-stained value in virus-transfected neurons. Scale bar, 200 μm/100 μm. K , Dendritic spine density of pyramidal neurons in mPFC. Scale bar, 10 μm. L , Levels of MAPK-related signal proteins, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ACE-CCtrl, ACE-C mice injected with Ctrl virus; ACE-COE, ACE-C mice with overexpression virus of DUSP1; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ACE-CCtrl mice.

Article Snippet: The transferred membranes were blocked with 5% non-fat dry milk and 0.1% Tween 20 in 10 mM Tris–HCl (TBST buffer, pH 7.5) for 1.5 h at room temperature, then subsequently incubated with the following primary antibodies: c-Fos (1:1500, Rabbit, RRID: AB_2247211, Cell Signalling Technology, USA), DUSP1 (1:1000, Rabbit, BM4856, Boster Biotechnology, China), ERK1/2 (1:1000, Rabbit, BM4326, Boster Biotechnology, China), p-ERK1/2 (1:1000, Rabbit, BM4156, Boster Biotechnology, China), CREB (1:1000, Rabbit, RRID:AB_331277, Cell Signaling Technology, USA), p-CREB (1:1000, Rabbit, RRID:AB_2561044, Cell Signaling Technology, USA), BDNF (1:1000, Mouse, RRID:AB_2881675, Proteintech, USA).

Techniques: Injection, Transfection, Activity Assay, Staining, Virus, Over Expression

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Experimental design and timeline. B , Schematic diagram of viral injection. Scale bar, 400 μm. C , Percentage of viral transfected neurons in CaMKII-positive neurons of mPFC. Scale bar, 200 μm/100 μm. D , Dusp1 mRNA levels on P78. E , DUSP1 protein levels on P78. F , DUSP1 activity on P78. G , CPP score. H , ΔCPP score. I , Heatmap of spent duration by mice in CPP apparatus. J , C-Fos-stained value in virus-transfected neurons. Scale bar, 200 μm/100 μm. K , Dendritic spine density of pyramidal neurons in mPFC. Scale bar, 10 μm. L , Levels of MAPK-related signal proteins, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ACE-CCtrl, ACE-C mice injected with Ctrl virus; ACE-CKD, ACE-C mice with knockdown virus of DUSP1; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ACE-CCtrl mice.

Article Snippet: The transferred membranes were blocked with 5% non-fat dry milk and 0.1% Tween 20 in 10 mM Tris–HCl (TBST buffer, pH 7.5) for 1.5 h at room temperature, then subsequently incubated with the following primary antibodies: c-Fos (1:1500, Rabbit, RRID: AB_2247211, Cell Signalling Technology, USA), DUSP1 (1:1000, Rabbit, BM4856, Boster Biotechnology, China), ERK1/2 (1:1000, Rabbit, BM4326, Boster Biotechnology, China), p-ERK1/2 (1:1000, Rabbit, BM4156, Boster Biotechnology, China), CREB (1:1000, Rabbit, RRID:AB_331277, Cell Signaling Technology, USA), p-CREB (1:1000, Rabbit, RRID:AB_2561044, Cell Signaling Technology, USA), BDNF (1:1000, Mouse, RRID:AB_2881675, Proteintech, USA).

Techniques: Injection, Transfection, Activity Assay, Staining, Virus, Knockdown

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: In adulthood, mice with cocaine-exposed experience during adolescent period (ACE-C mice) exhibits higher sensitivity to cocaine, accompanied by more activated mPFC and lower levels of DUSP1 protein and activity of mPFC in response to subthreshold-dosed administration of cocaine. Specific overexpression of DUSP1 on mPFC glutamatergic neurons efficiently blocks cocaine-preferred behaviors and reduces mPFC activity, while knockdown of DUSP1 maintains cocaine-preferred behaviors and increases mPFC activity in ACE-C mice. It might be through regulating MAPK signal molecules such as ERK1/2 that DUSP1 regulates the enhanced mPFC activation and sensitivity of ACE mice to cocaine.

Article Snippet: The transferred membranes were blocked with 5% non-fat dry milk and 0.1% Tween 20 in 10 mM Tris–HCl (TBST buffer, pH 7.5) for 1.5 h at room temperature, then subsequently incubated with the following primary antibodies: c-Fos (1:1500, Rabbit, RRID: AB_2247211, Cell Signalling Technology, USA), DUSP1 (1:1000, Rabbit, BM4856, Boster Biotechnology, China), ERK1/2 (1:1000, Rabbit, BM4326, Boster Biotechnology, China), p-ERK1/2 (1:1000, Rabbit, BM4156, Boster Biotechnology, China), CREB (1:1000, Rabbit, RRID:AB_331277, Cell Signaling Technology, USA), p-CREB (1:1000, Rabbit, RRID:AB_2561044, Cell Signaling Technology, USA), BDNF (1:1000, Mouse, RRID:AB_2881675, Proteintech, USA).

Techniques: Activity Assay, Over Expression, Knockdown, Activation Assay

Journal: Medicine

Article Title: Identification of disease-specific genes related to immune infiltration in nonalcoholic steatohepatitis using machine learning algorithms

doi: 10.1097/MD.0000000000038001

Figure Lengend Snippet: Correlation between NASH disease signature genes and immune cells. (A) correlation of FOS with immune cells; (B) correlation of IGFBP2 with immune cells; (C) correlation of DUSP1 with immune cells; (D) correlation of IKZF3 with immune cells. NASH = nonalcoholic steatohepatitis.

Article Snippet: The membrane was blocked with 5% skim milk for 2 hours and then incubated overnight at 4°C with primary antibodies against FOS (1:1000, cat. no. A17351; ABclonal), IGFBP2 (1:1000, cat. no. A0403; ABclonal), dual-specificity phosphatase 1 (DUSP1) (1:1000, cat. no. A22201; ABclonal), and IKZF3 (1:1000, cat. no. A8614; ABclonal).

Techniques:

Journal: Medicine

Article Title: Identification of disease-specific genes related to immune infiltration in nonalcoholic steatohepatitis using machine learning algorithms

doi: 10.1097/MD.0000000000038001

Figure Lengend Snippet: Protein expression in FOS, IGFBP2, DUSP1 and IKZF3 in HepG2 cells. (A) Protein bar graph; (B–E) protein expression statistics, compared with control, ** P < .01, *** P < .001, n = 3.

Article Snippet: The membrane was blocked with 5% skim milk for 2 hours and then incubated overnight at 4°C with primary antibodies against FOS (1:1000, cat. no. A17351; ABclonal), IGFBP2 (1:1000, cat. no. A0403; ABclonal), dual-specificity phosphatase 1 (DUSP1) (1:1000, cat. no. A22201; ABclonal), and IKZF3 (1:1000, cat. no. A8614; ABclonal).

Techniques: Expressing, Control