Figure 6 | Scientific Reports

Figure 6

From: Comparative analysis of lysyl oxidase (like) family members in pulmonary fibrosis

Figure 6

LOX/L knockdown leads to decreased smooth muscle actin production and stress fibre formation in NHLFs. (a) Primary human lung fibroblasts (NHLFs) were transfected with 16.6 nM of control, LOX, LOXL1, LOXL2, LOXL3 or LOXL4 siRNA. After 24 h, cells were stimulated with TGF-β (5 ng/ml) for 48 hours. Gene expression changes of acta2 were determined using Taqman gene expression assays and normalized to RNA-polymerase 2 expression and control treatment. The data represent the mean ± SD of three independent experiments (**p < 0.01; ***p < 0.001). (b) NHLFs were transfected as described in (a) and stimulated with TGF-β1 for 72 hours. αSMA fibrils per cell were quantified after immunocytochemical staining using high-content cellular imaging of 16 wells of a 384-well plate per condition (see “Image analysis” in the methods section for details). The data represent the mean ± SD of three independent experiments (**p < 0.01). Representative pictures of the cells stained for αSMA (red) and nuclei (DAPI, blue) are shown in (c). (d) acta2 induction by TGFβ1 on RNA level is decreased after FAK signaling pathway depletion. NHLF were transfected with 16.6 nM of control, ITGB3, FAK, AKT or SRF siRNA. After 24 h cells were stimulated with TGF-β1 (5 ng/ml) for 48 hours. Gene expression changes of acta2 were determined using Taqman gene expression assays. Data was normalized to RNA-polymerase 2 expression and control treatment and is presented as mean ± SD of three independent experiments (*p < 0.05).

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