POSTN Antibody - #DF6746

Figure 4. VGF activates CAFs to support SCLC proliferation and survival in the TME, A, RT-qPCR analysis of mRNA levels of CAF activation markers in MRC-5 cells after coculture with CM from indicated SCLC cells. B, Western blot analysis of αSMA and POSTN protein levels, two classic markers of CAF activation, in MRC-5 cells after coculture with CM from indicated SCLC cells. C–E, RT-qPCR analysis of mRNA levels of CAF activation markers in MRC-5 cells after coculture with CM from SCLC cells with VGF knockdown (C and D) or VGF overexpression (E). F, Western blot analysis of αSMA and POSTN protein levels in MRC-5 cells after coculture with CM from SCLC cells with VGF knockdown or overexpression. G and H, Immunofluorescence (G) and quantitative analysis (H) of αSMA expression and fibrillation in MRC-5 cells after coculture with CM from SCLC cells with VGF knockdown or overexpression. Scale bar, 5 μm. I and J, Wound healing and transwell assays to assess migration and invasion of MRC-5 cells after coculture with CM from SCLC cells with VGF knockdown or overexpression. Scale bar, 1 mm. K, Western blot analysis of TGFBR2, pSMAD2/3, and SMAD2/3 protein levels in MRC-5 cells following VGF overexpression or treatment with rVGF. L, CCK-8 assay measuring cell viability of SCLC cells after coculture with human primary isolated CAFs activated by CM from indicated SCLC cells. M, CCK-8 assay measuring cell viability of SCLC cells after coculture with human primary isolated CAFs activated by CM from SCLC cells with VGF knockdown or overexpression. Differences between groups were assessed using the unpaired t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3. Data are presented as the means ± SEM.

Fig. 3. Increased expression of periostin/nuclear factor kappa-B/C-X-C motif chemokine ligand 8 (POSTN/NF-κB/CXCL8) in long-term PD patients with peritoneal fibrosis. (a) Heatmap showing differential gene expression in PMCs from 1W-PD and 1Y-PD patients. (b) Representative immunofluorescence images of staining for POSTN, NF-κB P65, p-P65, CXCL8, α-smooth muscle actin (α-SMA), collagen I (Col-1), transforming growth factor-β (TGF-β), and FN in PMCs from 1W-PD and 1Y-PD patients. Scale bar = 50 µm. (c–i) mRNA expression levels in the two groups. (j–o) Representative Western blot images and quantitative analysis of POSTN, p-P65/P65, CXCL8, α-SMA, and Col-1 expression in the two groups.

Figure 2 Western blot analysis of periostin protein expression in HK-2 cells stimulated by HG (22 mmol/L) and high concentration of TGF-β1 (10 ng/mL); Image J software was used to collect the gray value of protein bands. *p