Figure 1
From: Endogenous HIF2A reporter systems for high-throughput functional screening
Generation of endogenous HIF2A reporter systems. (a) TCGA data analysis of HIF2A mRNA expression in ccRCC compared to different tumour types and normal kidney. P-value by Wilcoxon rank sum test. (b) Schematic of CRISPR-Cas9-based knock-in strategy of an mCherry fluorescent gene into the exon 16 of EPAS1. Plasmid template consists of an mCherry fluorescent gene with a hygromycin selection marker that are flanked by homology arms (HA). Sequencing primers used for genomic and cDNA amplifications in (c) and (d), respectively, are shown. (c) Genomic amplification of the HIF2A-mCherry integration site in single-cell derived clones. WC, water control. GC, genomic control. (d) cDNA amplification of the HIF2A-mCherry integration site. WC, water control. GC, genomic control. (e) Sanger sequencing of the HIF2A-mCherry integration sites in H2AmC2 cells. (f) FACS analysis of mCherry fluorescence in the three H2AmC clones compared to the parental UOK101 cells. (g) mCherry fluorescence in the three H2AmC clones compared to the parental control. DAPI in blue, mCherry in red.
