Fig. 2
From: Unbiased proteomic mapping of the LINE-1 promoter using CRISPR Cas9
C-BERST identification of 5’UTR biotinylated transcription factors. A Western blot of LINE-1 ORF1p expression in whole cell lysates of E006AA-hT and LNCaP cells. (Tubulin loading control). B Relative LINE-1 mRNA levels as assessed by qPCR. C Western blot of C-BERST streptavidin immunoprecipitation. Cells expressing gRNA-4, gRNA-7, or gRNA-NS were harvested, and biotinylated proteins were collected by streptavidin immunoprecipitation. A no hydrogen peroxide control (No H2O2) was included in the harvest to eliminate endogenously biotinylated proteins. Input, flow through, and immunoprecipitation (IP) were blotted with streptavidin HRP. D Proteins identified with C-BERST that were enriched at least 1.5 × in both LINE-1 targeting gRNAs (gRNA-4, gRNA-7) when compared to the non-targeting control (gRNA NS). Transcription factors have been highlighted in red. E Complete list of transcription factors enriched at least 1.5 × in both LINE-1 targeting gRNAs (gRNA-4, gRNA-7) when compared to non-targeting control (gRNA NS). Proteins previously identified to regulate LINE-1 are highlighted in green. Blue squares denote ENCODE ChIP data availability. Red squares denote peak on specified LINE-1 location by MapRRCon analysis