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. Author manuscript; available in PMC: 2020 Nov 4.
Published in final edited form as: Nat Struct Mol Biol. 2020 May 4;27(5):500–510. doi: 10.1038/s41594-020-0406-8

Figure 2. FLASH recruits CDK11 to the RDH genes.

Figure 2

a, Western blot analyses of immunoprecipitates of endogenous FLASH from HCT116 cells. The blots were probed with the indicated antibodies.

b, Depiction of human FLASH protein and four his-tagged deletion mutants expressed in bacteria. Deletion mutants A and B have an overlapping region between amino acids 490-571.

c, Western blot analyses of in vitro binding assays of GST-CDK11 purified from insect cells and his-tagged FLASH (HIS-FLASH) deletion mutants expressed in bacteria and depicted in Figure 2b.

d, FLASH ChIP-seq in hTERT cells (GSE69149) 36 (left panel) in comparison to CDK11 ChIP-seq (middle panel) and no Ab input control (right panel) on 44 regulated (expressed) RDH (RDH with base mean expression>10), all RDH and 200 other downregulated genes.

e, Endogenous CDK11 ChIP-qPCR on indicated RDH genes or control intergenic region (Ir) in HCT116 cells treated either with control (CTRL) or FLASH siRNAs for 24 h. n=3 biologically independent experiments, error bars=SEM, *P<0.05, Student´s two-sided t-test. Source Data for graphs in panel e are available with the paper on line.