Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: MMP28 recruits M2-type tumor-associated macrophages through MAPK/JNK signaling pathway-dependent cytokine secretion to promote the malignant progression of pancreatic cancer

doi: 10.1186/s13046-025-03321-x

Figure Lengend Snippet: MMP28 promotes the secretion of IL-8 and VEGFA from cancer cells by regulating the MAPK/JNK signaling pathway. A - B WB analysis of JNK, p-JNK, ERK, p-ERK, P38, and p-P38 in pancreatic cancer cells with MMP28 knockdown or overexpression compared with the control. C - D GEPIA database analysis of the relationships between JNK expression and the expression of IL-8 and VEGFA in pancreatic cancer. E ELISAs were used to measure IL-8 and VEGFA protein secretion in empty vector-treated pancreatic cancer cells, MMP28-overexpressing pancreatic cancer cells, and pancreatic cancer cells treated with the JNK inhibitor SP600125 (20 μM). **** P < 0.0001. ### P < 0.01, and #### P < 0.0001

Article Snippet: The membranes were blocked with 5% bovine serum albumin (GC305010-5 g, Servicebio) for one hour and incubated overnight at 4°C with primary antibodies against MMP28 (1:500, sc-515010, Santa Cruz Biotechnology), GAPDH (1:1000, GB12002, Servicebio), β-actin (1:1000, GB15003-100, Servicebio), JNK (1:500, BM4329, BOSTER), p-JNK (1:500 BM4380, BOSTER), P38-MAPK (1:500, BM4439, BOSTER), p-P38-MAPK (1:500, M00176T180, BOSTER), ERK (1:500, BM3426, BOSTER), p-ERK (1:500, BM4156, BOSTER), ANXA2 (1:1000, 11256-1-AP, Proteintech), Vimentin (1:500, PB9359, BOSTER), CDH1 (1:500, BM4166, BOSTER), CDH2 (1:500, BA0673, BOSTER), BAX (1:500, BA0315-2, BOSTER), and BCL-2 (1:500, A00040-2, BOSTER).

Techniques: Knockdown, Over Expression, Control, Expressing, Plasmid Preparation

Journal: PLoS ONE

Article Title: Endogenous Semaphorin-7A Impedes Human Lung Fibroblast Differentiation

doi: 10.1371/journal.pone.0170207

Figure Lengend Snippet: HLF derived from non-fibrotic lungs were treated with control-siRNA (Ctrl.) or semaphorin7A-siRNA (sema7A) for 24 h before starvation in BSA for 24 h. A/ Then cells were activated with 1% or 10% FBS for 15 min. ERK phosphorylation (P-ERK) was analyzed by western-blot. The graph shows ratios of phospho-ERK versus total ERK1, and is an average of 3 experiments. * indicates that the increase of phospho-ERK after semaphorin-7A (Sema7A)-siRNA-treatment is statistically different from phospho-ERK after control-siRNA (Ctrl) treatment. B/ Cells were activated with 1% FBS for 30 min and a representative blot of 2 experiments shows phosphorylation of p38 (P-p38). C/ Cells were treated with the indicated siRNA and cultured for 48 h in 1% FBS along with a phospho-ERK inhibitor (U0126) or its inactive analog (U0124) during the last 24 h. Proliferation was determined by adding BrdU in the last 6 h of the culture. One representative experiment of 2 using 2 different HLF lines is shown. Average ±SEM of 4 wells is shown.

Article Snippet: The rabbit anti-phospho-p38 antibody was from Genetel Laboratories (Madison, WI).

Techniques: Derivative Assay, Control, Phospho-proteomics, Western Blot, Cell Culture

Journal: Oncotarget

Article Title: Klotho inhibits EGF-induced cell migration in Caki-1 cells through inactivation of EGFR and p38 MAPK signaling pathways

doi: 10.18632/oncotarget.25481

Figure Lengend Snippet: (A) Cells were pretreated with EGF (100ng/ml) and harvested after the indicated times. Lysates were collected, and p38 MAPK phosphorylation levels were analyzed by Western blotting using the antiphospho-p38 antibody. The same membrane was striped and re-probed with a different antibody for measuring total-p38 MAPK protein levels. (B) Caki-1 cells were preincubated with 400pM of the secreted form of the Klotho (KL) protein at the indicated times followed by EGF (100ng/ml) stimulation for additional 5 min. Cell lysates were subjected to Western blot analysis as described in (A). Plots indicate means ± S.E.M of three independent experiments.

Article Snippet: Rabbit anti-human polyclonal active p38 MAPK antibody and SB203580 were purchased from Promega (Madison, WI).

Techniques: Phospho-proteomics, Western Blot, Membrane

Journal: Oncotarget

Article Title: Klotho inhibits EGF-induced cell migration in Caki-1 cells through inactivation of EGFR and p38 MAPK signaling pathways

doi: 10.18632/oncotarget.25481

Figure Lengend Snippet: (A) Classical in vitro wound healing assay was performed using near-confluent serum-starved Caki-1 cells grown on collagen type 1. Cells were either pretreated with 400pM Klotho (KL) or 1μM p38 MAPK-specific inhibitor SB203580 as positive control, for 60 min followed by EGF (100ng/ml) treatment. Cell culture images shown here were taken at 0 and 24 h. (B) Plots of quantification of the resultant cell motility values were computed by gap surface area measurements for four selected microscopic fields in each assay condition. The degree of migration is expressed as % wound closure compared with zero time point. The results represent means ± S.E.M of three independent experiments. (C) Wound healing assays performed under 3D settings. Images are microphotographs of Caki-1 cells showing hole-closure of “tissue openings” generated with magnetic pattering as described in the materials and method. Cells were pretreated the same way with either Klotho (KL) or SB203580 and stimulated with EGF as described for the classical wound healing assay. (D) Plots of rate closure of holes for Caki-1 cells as a function of EGF exposure in the presence or absence of KL or SB203580.

Article Snippet: Rabbit anti-human polyclonal active p38 MAPK antibody and SB203580 were purchased from Promega (Madison, WI).

Techniques: In Vitro, Wound Healing Assay, Positive Control, Cell Culture, Migration, Generated

Journal: Molecular Medicine Reports

Article Title: Placental growth factor gene silencing mitigates the epithelial-to-mesenchymal transition via the p38 MAPK pathway in rats with hyperoxia-induced lung injury

doi: 10.3892/mmr.2019.10785

Figure Lengend Snippet: Expression of p38 and p-p38 MAPK in hyperoxia-exposed lung tissue. Neonatal rats were exposed to normoxic or hyperoxic conditions for 14 days; hyperoxia exposed rats were subsequently injected with shRNA-NC or shRNA-PLGF. (A) Western blot analysis and (B) quantification was performed to determine p-p38/p38 protein expression levels in rat lung tissues; β-actin was used as the endogenous control. Data are presented as the mean ± standard deviation. n=8. ***P<0.001. MAPK, mitogen-activated protein kinase; NC, negative control; PLGF, placental growth factor; shRNA, short hairpin RNA.

Article Snippet: Anti-p38 rabbit monoclonal (cat. no. M00176), anti-β-actin goat polyclonal (cat. no. BM0627) and anti-ZEB2 rabbit polyclonal (cat. no. PA1959) antibodies were purchased from Boster Biological Technology.

Techniques: Expressing, Injection, shRNA, Western Blot, Control, Standard Deviation, Negative Control