Fig. 7
a-b RIP -qPCR assay of KYSE-150 cells was performed using an anti-PUM1 or control IgG antibody to detect NORAD expression (a) and pri-miR-199a1 expression (b). Values were normalized to the levels immunoprecipitated with the normal control IgG. c PUM1 protein expression was measured by using western blot in KYSE-150 and TE-1 cells transfected with SH-NORAD or SH-NC. d Effects of 0, 4 Gy, 6 Gy, and 8 Gy of X-ray radiation treatment on PUM1 protein expression were measured by using western blotting in KYSE-150 and TE-1 cells. e qRT–PCR assay to evaluate the expression of pri-miR-199a1 in KYSE-150 cells transduced with si-PUM1 or si-NC. f qRT–PCR assay to evaluate the expression of miR-199a-5p in KYSE-150 cells transduced with si-PUM1 or si-NC. g qRT–PCR assay to evaluate the expression of pri-miR-199a1 in NORAD knockdown cells and NORAD knockdown cells transduced with si-PUM1. h The effect of PUM1 or PUM1-ΔHD overexpression on pri-miR-199a1 expression in KYSE-150 cells treated with or without 8 Gy of X-ray radiation. (n = 3 technical replicates, *P < 0.05, **P < 0.01, *** P < 0.001, ****P < 0.0001, qRT–PCR results for miRNAs were quantified relative to GAPDH)