Fig 2: siRNA-mediated silencing of CDH2, LEP, POSTN, TIMP1 and VEGFC increased the proportion of PI-positive colon cancer cells. After 5-FU treatment (1 µg/ml) and silencing of CDH2, LEP, POSTN, TIMP1 or VEGFC, PI-positive (A) HCT116 and (B) LoVo cells were measured. The data was analyzed using an unpaired Student's t-test and expressed as the mean ± SD. Each assay was performed independent 3 times. ***P<0.001. si, small interfering; 5-FU, 5-fluorouracil.

Fig 3: Effect of ANGPTL8 on the migration and invasion of trophoblast cells. (A) Wound healing assay and (B) Transwell assay were performed to investigate the effect of ANGPTL8 on cell migration and invasion, respectively (magnification, x100). (C) The effect of ANGPTL8 on the protein expression levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 was analyzed by western blotting. The data represent the mean ± standard error of the mean of three independent experiments. ***P<0.001 vs. control or si-NC group. ANGPTL8, angiopoietin-like protein 8; si-, small interfering RNA; NC, negative control; MMP, matrix metalloproteinase; TIMP, tissue inhibitor of matrix metalloproteinase.

Fig 4: Modulation of the expression of MMPs/collagen in UV-B-irradiated NHDFs by NK-CdM. After being serum-starved for 24 h, NHDFs were irradiated with UV-B (30 mJ/cm2), and further incubated with NK-CdM at the indicated concentrations for 48 h. (A) The expression levels of procollagen I, type I collagen, type III collagen, a-elastin and (B) a quantitative densitometric analysis of the upper bands. (C) MMP-1, -2, -3, -9, and TIMP-1 were evaluated by western blotting. (D) The bar graph (mean ± standard deviation of the mean; n=3) represent a quantitative densitometric analysis of the upper bands. Changes in the mRNA expression of (E) type I collagen and (F) type III collagen were determined using quantitative PCR at 0, 6, 12, 24 and 48 h following culture in NK-CdM after UV-B exposure. The expression of photoaging-related genes such as (G) MMP-1, (H) MMP-3, (I) MMP-9, and (J) TIMP-1 was evaluated by quantitative PCR. Equal protein and mRNA loading were verified by normalization with ß-actin and GAPDH, respectively. *P<0.05, **P<0.01 and ***P<0.001 vs. with the UV-B-irradiated control. #P<0.05, ##P<0.01 and ###P<0.001 vs. the Normal (non-irradiated control). MMP, matrix metalloproteinase; NK-CdM, natural killer cell conditioned medium; NHDF, neonatal human dermal fibroblasts; UV, ultraviolet; TGF, transforming growth factor; TIMP, tissue inhibitor of matrix metalloproteinases.

Fig 5: C7A suppresses STAT3 phosphorylation and translocation in TGF-ß1-activated LX-2 cells. LX-2 cells were treated with C7A for 48 h after TGF-ß1 induction for 48 h. (A) The expression levels of p-STAT3 and STAT3 were analyzed by western blot assay. (B) The expression levels of p-STAT3 were analyzed from nuclear and cytosolic protein fractions. Lamin A/C was used as a nuclear loading control. (C) p-STAT3 (green) localization in LX-2 cells was determined by confocal immunocytochemistry. The nuclei are counterstained with 4',6-diamidino-2-phenylindole (DAPI) (Blue). The scale bars represent 50 µm. (D) The protein expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured by western blot assay. Densitometric analysis is expressed as mean ± SD intensity of optical density obtained by three independent experiments. (E) The mRNA expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were evaluated by qPCR analysis. Each experiment was repeated three times, and values represent mean ± S.D. ## p < 0.01 compared with control, ** p < 0.01, * p < 0.05 compared with TGF-ß1 treatment group.