Amplification of TGFβ Induced ITGB6 Gene Transcription May Promote Pulmonary Fibrosis - Fig 2

<p><b>A.</b> iHBECs were treated with 10 μg/ml anti-TGFβ and <i>ITGB6</i> mRNA was measured by QPCR at increasing time points. Data are expressed as mean expression relative to control (0 h) ± SEM from three independent experiments. * p < 0.05 <b>B.</b> iHBECs were treated with 10 μM Alk5 inhibitor and <i>ITGB6</i> mRNA was measured by QPCR at increasing time points. Data are expressed as mean expression relative to control (0 h) ± SEM from three independent experiments. * p < 0.05 <b>C.</b> iHBECs were treated with 10 μg/ml anti-TGFβ or an isotype control and αvβ6 cell surface expression was measured by flow cytometry after 3 days. Three independent experiments were performed and histogram shows representative data from one experimental repeat. Grey = negative control, black solid = 0 μg/ml, black dotted = 10 μg/ml. <b>D.</b> Amalgamated data from three independent experimental repeats of the experiment described in Fig 2C are demonstrated as a bar chart showing mean fold change in MFI ± SEM. * p < 0.05 <b>E.</b> iHBECs were treated with 0 or 10 μM Alk5 inhibitor (SB431542) and αvβ6 cell surface expression was measured by flow cytometry after 3 days. Three independent experiments were performed and histogram shows representative data from one experimental repeat. Grey = negative control, black solid = 0 μM, black dotted = 10 μM. <b>F.</b> Amalgamated data from three independent experimental repeats of the experiment described in Fig 2E are demonstrated as a bar chart showing mean fold change in MFI ± SEM. ** p < 0.01 <b>G.</b> iHBECs were treated with 0 or 10 μg/ml anti-αvβ6 and <i>ITGB6</i> mRNA was measured by QPCR at increasing time points. Data are expressed as mean expression relative to control (0 h) ± SEM from three independent experiments. ** p < 0.01, *** p < 0.005 <b>H.</b> iHBECs transfected with either pGL3-<i>ITGB6</i> or empty pGL3 as a control were treated with 0 or 10 μg/ml anti-αvβ6 for 4 hours. Data are expressed as mean relative firefly / renilla luciferase ratio ± SEM from three independent experiments. * p < 0.05 <b>I.</b> iHBECs were pre-treated with no antibody (black bars), 10 μg/ml anti-TGFβ (checked bars) or 10 μg/ml anti-αvβ6 (white bars) for one hour then stimulated with 20 μM LPA. <i>ITGB6</i> mRNA was measured by QPCR after 0 and 8 hours. Data are expressed as mean expression relative to control (0 h, no antibody) ± SEM from three independent experiments. * p < 0.05, ** p < 0.01 <b>J.</b> iHBECs were treated with 20μM LPA and <i>PAI</i> mRNA was measured by QPCR at 0, 2, 4 and 8 hours. Data are expressed as mean expression relative to control (0 h) ± SEM from three independent experiments. * p < 0.05.