Fig. 2

CBFB binds to RUNX1 mRNA via hnRNPK. a Silver staining of FLAG pulldown using CBFB KO MCF10A cells expressing empty vector and N-terminally FLAG tagged CBFB. b IB validation of CBFB and hnRNPK interaction. c IB showing the effect of hnRNPK knockdown on RUNX1 protein. d RNA immunoprecipitation (RIP) with CBFB antibody in WT and CBFB_KO MCF10A cells. Error bars are SEM, n = 3 (biological); two asterisks, p value < 0.01. (RIP of CBFB in WT vs. in CBFB KO cells, two-tailed t test). e RIP with hnRNPK antibody. Error bars are SEM, n = 3 (biological); two asterisks, p value < 0.01 (RIP of hnRNPK in WT vs. CBFB KO cells, two-tailed t test). f RNA pulldown assays (RPA) determining the hnRNPK-bound region within RUNX1 mRNA. F1-4, fragment 1 to 4 of 3′ UTR of RUNX1 mRNA. See Methods for details. Numbers indicate the nucleotide positions. g Re-analyses of two public eCLIP datasets (GSM2423241 and GSM2423242) of hnRNPK on the RUNX1 locus. Nucleotide sequences of two poly-C tracts within the binding site of hnRNPK were shown. F3, fragment 3 of 3′-UTR; T14, truncation 14 of 3′-UTR F3; Mu1, mutation 1; Mu2, mutation 2. h RPA using recombinant CBFB in the absence or presence of recombinant hnRPNK. i RPA using recombinant hnRNPK in the absence or presence of recombinant CBFB