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. 2017 Feb 23;2(4):e91454. doi: 10.1172/jci.insight.91454

Figure 5. Hsp90 regulates αSMA and extracellular matrix gene expression.

Figure 5

Primary lung fibroblasts (CD45–Col1+) isolated from lung fibroblast cultures of TGF-α–transgenic mice on doxycycline (Dox) for 4 weeks and treated with vehicle or 17-AAG and control, Hsp90AA-specific, or Hsp90AB-specific siRNA. (A) Quantification of αSma, Fn1, Col1α and Col5α gene transcripts relative to Hprt in the lung-resident fibroblasts treated with vehicle or 17-AAG (1 μM) for 24 hours. Data are cumulative of 3 independent experiments with similar findings (n = 3/group). (B) Quantification of αSma, Fn1, Col1α, and Col5α gene transcripts relative to Hprt in the lung-resident fibroblasts treated with control, Hsp90AA-specific, or Hsp90AB-specific siRNA for 72 hours. Data are cumulative of 2 independent experiments with similar findings (n = 3–4/group). (C) Immunoblots for αSMA and FN1 in the lysates of lung fibroblasts treated with vehicle or 17-AAG (1 μM) for 48 hours. (D) Quantification of αSMA and FN1 proteins normalized to GAPDH levels in the total lysates of lung fibroblasts treated with vehicle or 17-AAG (1 μM) for 24 hours. Data are representative of 2 independent experiments with similar findings (n = 3/group). Data shown are mean ± SEM. An unpaired 2-tailed Student’s t test was performed to measure the significance. *P < 0.05, **P < 0.005, and ***P < 0.0005.

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