Cellular location and transcriptional effects of CPSF6-RARG fusion protein. (A) HeLa cells were transfected with pcDNA3.1 expression plasmids of vehicle (vector), myc-CPSF6, myc-RARG, myc-CPSF6-RARG-L, myc-CPSF6-RARG-S, and myc-PML-RARA, respectively. Immunofluorescence was performed with myc-tag antibody. Both CPSF6-RARG fusions were predominantly expressed in the nucleus. Original magnification ×630. (B) 293T cells were transfected with RARE Cignal reporter and pcDNA3.1 expression plasmids of vehicle (vector), RARG, CPSF6-RARG-L, CPSF6-RARG-S, PML-RARA, and RARA, respectively. Relative firefly luciferase expression of cell lysates was normalized to Renilla luciferase. The expression of vector control was set to 1. (C) 293T cells transfected with RARE Cignal reporter and the indicated constructs were treated with dimethyl sulfoxide (DMSO) or 1 μM ATRA for 48 hours. Relative firefly luciferase expression of cell lysates was normalized to Renilla luciferase. Ratios were normalized against the cells treated with DMSO. n = 4 separate experiments (A-C). All data are presented as mean ± SD.