Fig. 3: HPV18 E7-mediated TAZ expression in regulated by the ERK1/2-SP1 signalling axis.
From: The Hippo pathway transcription factors YAP and TAZ play HPV-type dependent roles in cervical cancer
A WWTR1 promoter schematic showing predicted SP1 binding sites. TSS = transcription start site. Made using BioRENDER.com. B WWTR1-promoter luciferase assay in HeLa cells following mithramycin A (Mith A) treatment (n = 3). C RT-qPCR analysis of WWTR1 and YAP1 expression in HeLa cells following Mith A treatment (n = 4). U6 was used as a loading control. D WWTR1-promoter luciferase assay in C33A cells following transfection of GFP-HPV18 E7 plus Mith A treatment (n = 3). E RT-qPCR analysis of WWTR1 expression in C33A cells following transfection of GFP-HPV18 E7 plus Mith A treatment (n = 4). U6 was used as a loading control. F Representative western blots from (E) probed for TAZ, GFP and GAPDH (n = 3). G Luciferase assay in HeLa cells with the WWTR1-promoter SP1 binding site mutants (n = 3). H Luciferase assay in C33A cells stably expressing HA-HPV18 E7 following transfection of the WWTR1-promoter SP1 binding site mutants (n = 3). I WWTR1-promoter luciferase assay in HeLa cells transfected with HPV18 E6/E7 targeting siRNA plus SP1 2TD overexpression (n = 3). J RT-qPCR analysis of WWTR1 expression in HeLa cells transfected with HPV18 E6/E7 targeting siRNA plus SP1 2TD overexpression (n = 3). U6 was used as a loading control. K WWTR1-promoter luciferase assay in C33A cells stably expressing HA-HPV18 E7 following U0126 treatment (n = 3). L RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing HA-HPV18 E7 following U0126 treatment (n = 3). U6 was used as a loading control. M Representative western blot of C33A cells stably expressing HA-HPV18 E7 following U0126 treatment probed for TAZ, pERK1/2 (T202/Y204), ERK1/2, HPV18 E7 and GAPDH (n = 4). N ChIP-qPCR analysis of SP1 binding to the WWTR1 promoter in HeLa and SiHa cells (n = 3). SP1 binding is presented as a fold increase over IgG binding and a gene desert control. Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file. Figure 3A was created with Biorender.com released under a Creative Commons Attribution-NonCommercial 4.0 International License.
