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. 2022 Sep 21;54(9):1549–1562. doi: 10.1038/s12276-022-00849-2

Fig. 3. Mechanical stress and TGF-β1 induce the expression of CLU in LF cells.

Fig. 3

a–c Quantitative RT‒PCR analysis showing the gene expression of TGF-β1, CLU and COL1A2 in LF cells after treatment for 8 h or 16 h under mechanical stress. Relative mRNA expression levels for nontreated cells were set as 1. Data are expressed as the mean ± S.D. *P < 0.05, **P < 0.01. d The nonstress group showed fibroblast‐like morphology with weak staining of TGF-β1 and CLU. The stress group showed a higher intensity of TGF-β1 and CLU staining after applying mechanical stress for 16 h. TGF-β1 (green), CLU (red), DAPI (blue); the scale bar indicates 50 μm. e–g ELISAs of TGF-β1, CLU and collagen I protein levels in culture medium after subjecting samples to mechanical stress for 8 h or 16 h. Data are shown as the mean ± S.D. *P < 0.05, **P < 0.01. h–j Quantitative RT‒PCR analysis showing the gene expression levels of CLU, α-SMA and COL1A2 in LF cells after TGF-β1 (5 ng/mL) treatment for the indicated duration. Relative mRNA expression levels for nontreated cells were set as 1. Data are expressed as the mean ± S.D. *P < 0.05, **P < 0.01. k Fibroblast‐like morphology with staining of CLU after treatment with 5 ng/mL TGF-β1 for the indicated duration showed an increase in staining intensity in a time-dependent manner. CLU (red), DAPI (blue); the scale bar indicates 50 μm. l Western blot analysis showing the protein levels of CLU, α-SMA and COL1A2 in LF cells after TGF-β1 (5 ng/mL) treatment for the indicated duration for the four groups. The expression levels of CLU, α-SMA and COL1A2 increased in a time-dependent manner.