Extended Data Fig. 5: nelfe-FKBP12 homozygous cell line generation.
From: Evolution of promoter-proximal pausing enabled a new layer of transcription control
a) Schematic of CRISPR design to add the FBBP12 tag at the nelfe locus. b) PCR validation of CRISPR insertion of the FKBP12 tag. c) Microscopy images evaluating the degradation efficiency before (top) and after a 30 min treatment with 500 nM dTAG-13 (bottom) in the edited and unedited cell lines. Hoechst was used as a nuclear control, while anti-HA antibodies measure the added tag, and anti-NELFE measures the NELF-E protein level. Arrows point out the presence of Feeder cells. d) Degradation efficiency of NELF-E as measured by western blotting. b-Actin was used as a loading control, while anti-HA measures the level of NELFE-HA protein. Input denotes the relative amount of total protein loaded.
