Journal: Molecular Medicine

Article Title: Metformin alleviates the calcification of aortic valve interstitial cells through activating the PI3K/AKT pathway in an AMPK dependent way

doi: 10.1186/s10020-021-00416-x

Figure Lengend Snippet: Metformin alleviates aortic calcification by activating the PI3K/AKT signaling pathway in a concentration-dependent manner in vitro. a Metformin inhibits inflammatory factor secretion including IL6, IL8, and MCP-1 in cell supernatant, after treatment with phosphate medium (PM) with or without metformin for 72 h as determined by ELISA; b ROS production was detected and quantified after PM treatment with or without metformin for 72 h; c Cell viability was detected by CCK8 after treatment with PM with or without metformin for 72 h; d Immunofluorescence staining images of p-AKT and OPN expression in AVICs after PM treatment with or without 100 μM metformin for 72 h, Scale bar, 50 μm; e The protein expression of p-AMPK (Thr172), p-AKT (Ser473), PI3K, p-eNOS (Ser1177), BMP2, and OPN in AVICs after PM treatment with or without metformin for 72 h as determined by WB; f Calcium deposition were detected by ARS Staining after various treatments for 7 days, Scale bar, 200 μm. n = 6 per group. CTL, control; Met, metformin; *p < 0.05 versus PM group (one-way ANOVA with Bonferroni post hoc test)

Article Snippet: The levels of IL6 (KE00007, Proteintech, China), IL8 (KE00006, Proteintech, China), and MCP-1 (KE00091, Proteintech, China) in the supernatant of AVICs were determined by ELISA.

Techniques: Concentration Assay, In Vitro, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Expressing, Control

Journal: Cell reports

Article Title: Collagen 1-mediated CXCL1 secretion in tumor cells activates fibroblasts to promote radioresistance of esophageal cancer.

doi: 10.1016/j.celrep.2023.113270

Figure Lengend Snippet: Figure 4. CXCL1 promotes migration and activation of fibroblasts via the CXCR2-STAT3 pathway (A) Western blot analysis of myCAF- and iCAF-related markers in fibroblasts incubated with CM from CXCL1-knockdown ESCC cells. (B) Quantification of migration in (Figure S5B) (n = 3 biological replicates). (C) Western blot analysis of myCAF- and iCAF-related markers in fibroblasts treated with indicated concentration of rCXCL1 proteins for 24 h. (D) Representative images of fibroblasts migration caused by indicated concentration of rCXCL1 proteins. Scale bar, 100 mm. (E) Quantification of migration in (D) (n = 3 biological replicates). (F) Western blot analysis of CAF markers in fibroblasts pre-treated with SB-265610 for 6 h and then treated by CM of KYSE510 for 18 h. KYSE510 cells were treated with Col1 or not. (G) Western blot analysis of CAF markers in fibroblasts pre-treated with SB-265610 for 6 h and then treated by rCXCL1 for 18 h. For all panels, data are presented as mean ± SD. **p < 0.01, ***p < 0.001, and ****p < 0.0001, as determined using two-tailed Student’s t test. Each assay for western blot had three biological repeats, and the quantification results are presented below each band. See also Figure S5.

Article Snippet: For CXCR2 receptor inhibitory assay, CXCR2 inhibitor SB-265610 (MCE, HY-50688) was added to serum-free medium for 6 h after fibroblasts grew until confluency, then CXCL1 recombinant protein was added to medium for another 18 h.

Techniques: Migration, Activation Assay, Western Blot, Incubation, Knockdown, Concentration Assay, Two Tailed Test

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: Anti-IL-8 and anti-VEGFA antibodies partially reverse the ability of MMP28 to promote the migration of TAMs and the polarization of M2 TAMs. A CM from MMP28-overexpressing pancreatic cancer cells was treated with an anti-IL-8 neutralizing antibody (5 μg/mL) or an anti-VEGFA neutralizing antibody (1 μg/mL). The migration level of TAMs was determined by the Transwell migration experiment. Scale bar, 100 μm. B qRT-PCR determination of CD86 and CD163 mRNA expression levels in TAMs cocultured with Vector group of pancreatic cancer cells transfected with empty vector virus, TAMs cocultured with OE-MMP28 cell group, TAMs cocultured with the OE-MMP28 + IgG cell group, TAMs cocultured with OE-MMP28 + Anti-IL-8 cell group, and TAMs cocultured with OE-MMP28 + Anti-VEGFA cell group. C The expression levels of the TAM markers CD86 and CD163 in different coculture groups were determined by immunofluorescence staining. Scale bar, 50 μm. D The proportions of CD11b + CD86 + TAMs and CD11b + CD163 + TAMs in different coculture groups were analysed by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: When the tumors reached a volume of at least 80 mm 3 one week post-injection, the treatment groups received the following treatments: PBS, Clodronate Liposomes (200 μL/mouse, YEASEN), the JNK inhibitor SP600125 (15 mg/kg/mouse, MCE), the interleukin-8 (IL-8)-neutralizing antibody MAB208-SP (10 μg/mouse, R&D Systems), or vascular endothelial growth factor A (VEGFA)-neutralizing antibody MAB293-SP (1 μg/mouse, R&D Systems).

Techniques: Migration, Quantitative RT-PCR, Expressing, Plasmid Preparation, Transfection, Virus, Immunofluorescence, Staining, Flow Cytometry

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 enhances M2 TAM infiltration and promotes pancreatic cancer growth in vivo. A Effects of the administration of Clodronate Liposomes, the JNK inhibitor SP600125, the anti-IL-8 neutralizing antibody MAB208-SP, and the anti-VEGFA neutralizing antibody MAB293-SP on subcutaneous tumors in nude mice in the experimental group. B - C Representative images of subcutaneous tumors in nude mice detected with fluorescein potassium salt and the fluorescence quantitative analysis. After the tumor volume reached at least 80mm 3 , PBS, Clodronate Liposomes (200 μg), the JNK inhibitor SP600125 (15 mg/kg), the anti-IL-8 antibody MAB208-SP (10 μg), and the anti-VEGFA antibody MAB293-SP (1 μg) were injected intraperitoneally into the nude mice. D Representative images of subcutaneous xenograft tumors (5 mice per group). E The volume of the tumor was measured every four days. F Tumor weight in both groups. G Representative images of MMP28 expression levels in xenograft tumors from the Vector and OE-MMP28 groups as determined by immunohistochemical staining. Scale bar, 50 μm. H Immunohistochemical staining detection of expression of Ki67 and CD86 + and CD163 + TAM infiltration in xenograft tumors in the Vector group, OE-MMP28 group, OE-MMP28 + SP600125 group, OE-MMP28 + MAB208-SP group, OE-MMP28 + MAB293-SP group, and Vector + Clodronate Liposomes group

Article Snippet: When the tumors reached a volume of at least 80 mm 3 one week post-injection, the treatment groups received the following treatments: PBS, Clodronate Liposomes (200 μL/mouse, YEASEN), the JNK inhibitor SP600125 (15 mg/kg/mouse, MCE), the interleukin-8 (IL-8)-neutralizing antibody MAB208-SP (10 μg/mouse, R&D Systems), or vascular endothelial growth factor A (VEGFA)-neutralizing antibody MAB293-SP (1 μg/mouse, R&D Systems).

Techniques: In Vivo, Liposomes, Fluorescence, Injection, Expressing, Plasmid Preparation, Immunohistochemical staining, Staining