Figure 6.

CRISPR-targeted knockdown of PTPRA results in attenuated responses of human fibroblasts to transforming growth factor-β (TGF-β) stimulation. A: CRISPR-targeted knockdown of PTPRA in hTERT-transformed immortalized human lung fibroblasts (IHLFs) results in efficient deletion of the protein tyrosine phosphatase-α (PTPα) protein by Western blotting. B: PTPRA-sufficient and PTPRA-KD fibroblasts were treated with 2 ng/mL of TGF-β, or buffer for 24 h and harvested to analyze the expression of SERPINE1, CTGF, and CYR61 by qPCR. C: PTPRA-sufficient and PTPRA-KD fibroblasts were treated with 2 ng/mL of TGF-β for 30 min and harvested, after which phosphorylation status of SMAD3 was assessed by Western blotting. Densitometry shows expression of phospho-SMAD3 over total SMAD3. D: PTPRA-sufficient and PTPRA-KD fibroblasts were treated with 2 ng/mL of TGF-β for 15 min and harvested, after which phosphorylation status of p38 was assessed by Western blotting. Densitometry shows expression of phospho-p38 over total p38. Data represent mean values ± SE from at least three independent experiments. *P < 0.05; one-way ANOVA.