anti-phosphorylated p38mapk  (Servicebio Inc)

Journal: Scientific Reports

Article Title: The hydroxamate based HDAC inhibitor WMJ-J-09 induces colorectal cancer cell death by targeting tubulin and downregulating survivin

doi: 10.1038/s41598-025-04714-w

Figure Lengend Snippet: LKB1-p38 MAPK signaling contributed to WMJ-J-09-mediated p21 and survivin regulation. ( A ) Immunoblot result of p38MAPK phosphorylation in HCT116 cells exposed to WMJ-J-09. ( B , C ) Immunoblot result of p21 ( B ) and survivin ( C ) expression in WMJ-J-09-stimulated HCT116 cells with or without p38 inhibitor III ( D ) Immunoblot result of LKB1 phosphorylation in HCT116 cells exposed to WMJ-J-09 for indicated periods. ( E ) Immunoblot results from the effects of LKB1 siRNA or negative control siRNA on p38MAPK and p53 phosphorylation elicited by WMJ-J-09. ( F ) Immunoblot result of the effects of LKB1 siRNA or negative control siRNA on WMJ-J-09-modulated p21 and survivin expression in HCT116 cells. Each band intensity was quantified, and total α-tubulin levels normalized the fold changes of LKB1, p21, and survivin; total p53 levels normalized that of p53 phosphorylation; total p38MAPK levels normalized that of p38MAPK phosphorylation; total LKB1 levels normalized that of LKB1 phosphorylation. Error bars, mean ± S.E.M. (shown only for independent replicate experiments with n ≥ 4). One-way ANOVA followed by Tukey’s post-hoc test assessed statistical significance (*p < 0.05 compared to the control group).

Article Snippet: Antibodies against α-tubulin Lys40 acetylation, p53, p53 Lys379 acetylation, p53 Ser15 phosphorylation, p38MAPK Thr180/Tyr182 phosphorylation, p38MAPK, LKB1, LKB1 Ser428 phosphorylation, PARP, cleaved caspase 3 (active form) and survivin were from Cell Signaling (Danvers, MA, U.S.A.).

Techniques: Western Blot, Phospho-proteomics, Expressing, Negative Control, Control

Journal: Scientific Reports

Article Title: The pro-tumor activity of INTS7 on lung adenocarcinoma via inhibiting immune infiltration and activating p38MAPK pathway

doi: 10.1038/s41598-024-77093-3

Figure Lengend Snippet: Potential mechanisms involved in the regulatory activity of INTS7. (A) Immunohistochemical staining and statistical analysis of INTS7 in clinical tumor tissues ( n = 6, Low = Group with low INTS7 expression, High = Group with high INTS7 expression, scale bar: 50 μm). (B) Immunohistochemical staining and statistical analysis of cleaved caspase-3 (c-caspase-3) in clinical tumor tissues ( n = 6, Low = Group with low INTS7 expression, High = Group with high INTS7 expression, scale bar: 50 μm). (C) H&E staining of xenograft tumor tissues (scale bar: 50 μm). (D) Immunohistochemical staining and statistical analysis of INTS7 in xenograft tumor tissues ( n = 5, scale bar: 50 μm). (E) Immunohistochemical staining and statistical analysis of cleaved caspase-3 (c-caspase-3) in xenograft tumor tissues ( n = 5, scale bar: 50 μm). (F) Expression of INTS7 by western blot analysis ( n = 3). (G) Expression of cleaved caspase-3 (c-caspase-3), p53, phosphorylated p38MAPK (p-p38MAPK) and MMP2 by western blot analysis ( n = 3, respectively). The grouping of blots were cropped from different parts of the same gel. Original images of blots are presented in Supplementary Figs. 2–6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) and incubated overnight at 4 ℃ with the following primary antibodies: anti-INTS7 (1:1000, Proteintech, USA), anti-cleaved caspase-3 (1:1000, Proteintech, USA), anti-p53 (1:500, Proteintech, USA), anti-p38MAPK (1:1000, Servicebio, China), anti-phosphorylated p38MAPK (1:1000, Servicebio, China), anti-MMP2 (1:1000, Servicebio, China). β-actin (1:5000, Proteintech, USA) was used as loading control.

Techniques: Activity Assay, Immunohistochemical staining, Staining, Expressing, Western Blot

Journal: Cells

Article Title: Mitophagy Effects of Protodioscin on Human Osteosarcoma Cells by Inhibition of p38MAPK Targeting NIX/LC3 Axis.

doi: 10.3390/cells12030395

Figure Lengend Snippet: Figure 5. The p38MAPK signaling pathway mediates the PD-induced mitophagy and apoptosis in human OS cells. (A,B) Human HOS and 143B OS cells were exposed to various concentrations (0, 2, 4, and 6 µM) of PD for 30 min. Cells were harvested and examined with immunoblotting to deter-mine the signaling pathway activation. HOS cells exposed to 4 µM PD and/or p38 siRNA (10 nM) were harvested for detecting (C) MMP changes, (D) AO staining, (E) mitophagy induction, and (F) the protein expressions of p38, cleaved caspase-3, cleaved PARP, NIX, and LC3B proteins, using immunoblotting assays. GAPDH was used as an internal loading control. ** p < 0.01, versus the control. # p < 0.05 versus the PD-only treatment (line 2). The results are the mean ± standard deviation of three experiments. Scale bar: 50 µm.

Article Snippet: Antibodies against cleaved caspase-caspase-3 (SC-56053), PINK1 (SC-518052), Parkin (SC-32282), NIX (SC-166332), BNIP3 (SC-56167), total p38MAPK (SC-7972), phosphoryl p38MAPK (SC-166182), total Akt (SC-56878), phosphoryl Akt (SC-271966), LC3 siRNA (SC-43390), NIX siRNA (SC-37453), and horseradish peroxidase-conjugated secondary antibodies (anti-mouse and anti-rabbit) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Western Blot, Activation Assay, Staining, Control, Standard Deviation