Figure 3 from KRAS Promotes GLI2-Dependent Transcription during Pancreatic Carcinogenesis

Abstract

<p>Gli2 is required for the regulation of <i>Ccnd1</i> downstream of oncogenic KRAS. <b>A,</b> Heatmap of GLI target gene expression in 1012U cells at 12, 24, 48, and 72 hours posttreatment with doxycycline (Dox). <b>B,</b> Relative gene expression of <i>Ccnd1</i> by qPCR in mouse pancreas tissue from Cre, CRG, KC, and KCRG mice. <b>C,</b> Western blot showing expression of p-ERK, total ERK, and CCND1 in 1012U −Dox and +Dox cells at 12, 24, 48, and 72 hours post treatment. Vinculin is used as the loading control. <b>D,</b> Relative gene expression of <i>Ccnd1</i> and <i>Gli2</i> by qPCR after the knockdown of <i>Gli2</i> by siRNA in 1012U cells +Dox cells (72 hours treatment; <i>P</i> < 0.05 and <i>P</i> < 0.001). <b>E,</b> Western blot (left) and protein quantification (right) representing expression of CCDN1 and GLI2 in 1012U cells +Dox, after the knockdown of <i>Gli2</i>. Vinculin is used as the loading control. <b>F,</b> Western blot (left) and protein quantification (right) representing expression of CCDN1 and GLI2 in 1012U cells +Dox and murine KC cell line transfected with <i>Gli2</i>, <i>∆NGli2</i>, or empty vector (control). Vinculin is used as the loading control. <b>G,</b> Top: Graphical representative of mouse <i>Ccnd1</i> promoter region with (red) diamond indicating candidate Gli2-binding site. Bottom: Graph representing enrichment (percent input) of GLI2 at the <i>Ccnd1</i> promoter in 1012U +Dox cells (72 hours treatment; <i>P</i> < 0.05).</p>