Fig. 4

In vitro recombinant human THBS2 protein promotes the collagen synthesis and invasive phenotype. (A) The mRNA expression levels of COL1A1 and COL3A1 in N-FBs stimulated with different concentrations of rhTHBS2 were detected by RT-qPCR. n = 6 for each group. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ^nsP > 0.05. (B) Western blot analysis of COL1A1, COL1A2, COL3A1, LOX and LOXL2 protein expression in N-FBs stimulated with different concentrations of rhTHBS2 for 48H, with the most significant increase at 200 ng/ml. n = 6 for each group. (C) Quantitative analysis of protein levels of COL1A1, COL1A2, COL3A1, LOX and LOXL2. n = 6 for each group. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ^nsP > 0.05. (D) Western blot analysis of COL1A1, COL1A2, COL3A1, LOX and LOXL2 protein expression at different time points after N-FBs stimulation with 200 ng/ml rhTHBS2, with the most significant increase at 48H. n = 6 for each group. (E) Quantitative analysis of protein levels of COL1A1, COL1A2, COL3A1, LOX and LOXL2. n = 6 for each group. Data are shown as the mean ± SEM. **P < 0.01, ****P < 0.0001, ^nsP > 0.05. (F) After stimulation of N-FBs with 200 ng/ml rhTHBS2 for 48 h, equal amounts of fibroblasts were loaded into transwell chambers. Images of migrating fibroblasts from the crystal violet staining assay are shown. Scale bar, 200 μm. n = 6 for each group. Data are shown as the mean ± SEM. **P < 0.01. (G) After stimulation of N-FBs with 200 ng/ml rhTHBS2 for 48 h, the proliferation of N-FBS was detected by EdU assay. Scale bar, 100 μm. n = 6 for each group. Data are shown as the mean ± SEM. ****P < 0.0001.