Journal: Molecular Oncology
Article Title: Identification of novel GLI1 target genes and regulatory circuits in human cancer cells
Figure Lengend Snippet: Reciprocal proliferation effects of GLI1 and FOXS1; FOXS1 inhibits GLI1 activity. EdU incorporation assay of Rh36 cells (A) and Daoy cells (B) following transfection with siRNA combinations of siControl + siControl, siControl + siGLI1, siControl + siFOXS1, or siGLI1 + siFOXS1. Data from one representative experiment are shown in the histogram. EdU incorporation assay of Daoy cells (C) following transfection/transduction with siControl + Ad‐Vector, siControl + Ad‐GLI1, siFOXS1 + Ad‐Vector, or siFOXS1 + Ad‐GLI1. Adenoviruses were added 6 h after siRNA transfection. Data from one representative experiment are shown in the histogram. (D) A schematic diagram of the proposed model for the FOXS1/GLI1 feedback loop. The transcription factor GLI1 positively regulates the expression of the FOXS1 transcription factor, while FOXS1 negatively regulates GLI1. GLI1 promotes cell proliferation, whereas FOXS1 inhibits cell proliferation. (E) FOXS1 expression reduces GLI1 transcriptional activity. HEK293A cells were co‐transfected with pcDNA3.1 vector (pcDNA), pGL3 basic luciferase empty vector (pGL3), pCMV‐GLI1‐flag (pGLI1), or pcDNA3.1‐FOXS1 (pFOXS1), together with the reporter plasmid 12xGLIBS and the control plasmid Renilla. Data from one representative experiment are shown. Error bars indicate standard deviation. Statistical significant, ## P
Article Snippet: HEK293A cells were seeded in 24‐well plates and transfected with 200 ng of 12xGLI binding site luciferase reporter plasmid (12xGLIBS) (Mao et al ., ) and 10 ng of Renilla control reporter plasmid, together with pcDNA3.1 vector, pGL3 basic luciferase empty vector, the pCMV‐GLI1‐flag expression construct (pGLI1), and/or pcDNA3.1‐FOXS1 ( ; GeneScript, Piscatway, NJ, USA) using Lipofectamine 3000 (Invitrogen).
Techniques: Activity Assay, Transfection, Transduction, Plasmid Preparation, Expressing, Luciferase, Standard Deviation