phosphorylated p38 mapk p p38  (Cell Signaling Technology Inc)

Journal: Molecular Medicine Reports

Article Title: PLEKHA4 knockdown induces apoptosis in melanoma cells through the MAPK and β‑catenin signaling pathways

doi: 10.3892/mmr.2025.13464

Figure Lengend Snippet: PLEKHA4 knockdown suppresses MAPK signaling. (A) Tandem mass tag proteomics was performed. Principal component analysis using the reporter ion log 2 ratios of all analyzed proteins showed that shPLEKHA4 A375 cells had a distinct proteomic profile compared with shNC cells. (B) Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that MAPK signaling was inhibited by PLEKHA4 knockdown. (C) Integrated functional association networks of PLEKHA4, β-catenin-related, MAPK-related and MYC genes. (D) Correlation analysis between PLEKHA4 and MAPK1. (E) Correlation analysis between PLEKHA4 and MAPK3. (F) Correlation analysis between PLEKHA4 and JUN. PLEKHA4, pleckstrin homology domain-containing family A member 4; sh, short hairpin; NC, negative control.

Article Snippet: The primary antibodies used included PLEKHA4 (cat. no. NBP1-56679; Novus Biologicals; Bio-Techne), β-actin (cat. no. GB12001-100; Wuhan Servicebio Technology), phosphorylated (p)-p38 MAPK (cat. no. AF4001; Affinity Biosciences), p38 MAPK (cat. no. AF6456; Affinity Biosciences), p-JNK1/2/3 (cat. no. AF3318; Affinity Biosciences), JNK1/2/3 (cat. no. AF6318; Affinity Biosciences), p-cJUN (cat. no. AF3095; Affinity Biosciences), c-Jun (cat. no. AF6090; Affinity Biosciences), p-MEK1/2 (cat. no. AF8035; Affinity Biosciences), MEK (cat. no. AF6385; Affinity Biosciences), p-ERK (cat. no. AF1015; Affinity Biosciences), ERK1/2 (cat. no. AF0155; Affinity Biosciences), COX2 (cat. no. AF7003; Affinity Biosciences), p-NF-κB p65 (cat. no. AF2006; Affinity Biosciences), NF-kB p65 (cat. no. AF5006; Affinity Biosciences), cleaved-caspase-3 (cat. no. AF2006; Affinity Biosciences), caspase-3 (cat. no. AF6311; Affinity Biosciences) cMyc (cat. no. 10828-1-AP; Proteintech Group), ubiquitin polyclonal antibody (cat. no. 10201-2-AP; Proteintech Group), β-catenin (cat. no. AF6266; Affinity Biosciences), p-GSK3β (cat. no. sc-373800; Santa Cruz Biotechnology), GSK3β (cat. no. sc-377213; Santa Cruz Biotechnology), cyclin D1 (cat. no. AF0931; Affinity Biosciences) and Lamin B1 (cat. no. sc-374015; Santa Cruz Biotechnology), each diluted 1:1,000 in Primary Antibody Dilution Buffer (Beyotime Institute of Biotechnology).

Techniques: Knockdown, Functional Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: PLEKHA4 knockdown induces apoptosis in melanoma cells through the MAPK and β‑catenin signaling pathways

doi: 10.3892/mmr.2025.13464

Figure Lengend Snippet: Impact of PLEKHA4 knockdown and overexpression on MAPK pathway protein expression in melanoma cells. (A) Western blotting of proteins in the MAPK pathway after PLEKHA4 knockdown **P<0.01, ***P<0.001vs. shNC. (B) Western blotting of proteins in the MAPK pathway after PLEKHA4 overexpression. # P<0.05, ### P<0.001 and #### P<0.0001 vs. NS. PLEKHA4, pleckstrin homology domain-containing family A member 4; sh, short hairpin; NC, negative control; NS, empty vector; p-, phosphorylated.

Article Snippet: The primary antibodies used included PLEKHA4 (cat. no. NBP1-56679; Novus Biologicals; Bio-Techne), β-actin (cat. no. GB12001-100; Wuhan Servicebio Technology), phosphorylated (p)-p38 MAPK (cat. no. AF4001; Affinity Biosciences), p38 MAPK (cat. no. AF6456; Affinity Biosciences), p-JNK1/2/3 (cat. no. AF3318; Affinity Biosciences), JNK1/2/3 (cat. no. AF6318; Affinity Biosciences), p-cJUN (cat. no. AF3095; Affinity Biosciences), c-Jun (cat. no. AF6090; Affinity Biosciences), p-MEK1/2 (cat. no. AF8035; Affinity Biosciences), MEK (cat. no. AF6385; Affinity Biosciences), p-ERK (cat. no. AF1015; Affinity Biosciences), ERK1/2 (cat. no. AF0155; Affinity Biosciences), COX2 (cat. no. AF7003; Affinity Biosciences), p-NF-κB p65 (cat. no. AF2006; Affinity Biosciences), NF-kB p65 (cat. no. AF5006; Affinity Biosciences), cleaved-caspase-3 (cat. no. AF2006; Affinity Biosciences), caspase-3 (cat. no. AF6311; Affinity Biosciences) cMyc (cat. no. 10828-1-AP; Proteintech Group), ubiquitin polyclonal antibody (cat. no. 10201-2-AP; Proteintech Group), β-catenin (cat. no. AF6266; Affinity Biosciences), p-GSK3β (cat. no. sc-373800; Santa Cruz Biotechnology), GSK3β (cat. no. sc-377213; Santa Cruz Biotechnology), cyclin D1 (cat. no. AF0931; Affinity Biosciences) and Lamin B1 (cat. no. sc-374015; Santa Cruz Biotechnology), each diluted 1:1,000 in Primary Antibody Dilution Buffer (Beyotime Institute of Biotechnology).

Techniques: Knockdown, Over Expression, Expressing, Western Blot, Negative Control, Plasmid Preparation

Journal: Molecules

Article Title: Anti-Inflammatory Effects of the Combined Treatment of Resveratrol- and Protopanaxadiol-Enriched Rice Seed Extract on Lipopolysaccharide-Stimulated RAW264.7 Cells

doi: 10.3390/molecules29184343

Figure Lengend Snippet: Effect of resveratrol- and PPD-enriched rice seed extracts (DJ526 and DJ-PPD, respectively) on the expression of immune-associated proteins in LPS-stimulated RAW264.7 cells. ( a ) Representative of blot analysis, ( b ) densitometric analyses of p-p38 MAPK protein expression, ( c ) densitometric analyses of p-ERK 1/2 protein expression, and ( d ) densitometric analyses of p-NF-κB p65 protein expression. The concentrations of DMSO, aspirin, and treatments (rice seed extract and mixture) were 0.1%, 200 ng/mL, and 100 µg/mL, respectively. The significant differences in protein expression are presented as lowercase letters at p < 0.05.

Article Snippet: Membranes were incubated with antibodies in blocking solution at the ratio 1:2000 ( v / v ) specific for phosphorylation (p)-p38 MAPK (Cell Signaling Technology, Danvers, MA, USA), p-NF-κB p65 (Cell Signaling Technology), and p-ERK 1/2 (Cell Signaling Technology).

Techniques: Expressing