Fig. 6
Expression and inhibition of RORC2 in ALCL. a Left, analysis of RORC1 and RORC2 mRNA by RT-PCR (top) and RORC protein expression by immunoblotting of nuclear extracts (bottom). GAPDH and PARP1 are controls. Right, AP-1 inhibition downregulates RORC2. K299 cell were treated as in Fig. 4d, and RORC2 and GAPDH mRNA were analyzed by RT-PCR (top), and RORC2 expression changes at the protein level by WB (bottom). Two (#1 and #2) of four independent experiments are shown. b RORC inhibition by small compounds induces cell death of ALCL cell lines. Cells were treated with 5 μM of RORC inhibitors SR2211 or SR1903, or the control (DMSO), and the percentage of viable cells was analyzed by PI staining. One of three independent experiments is shown. (c) Enforced cell death induction by crizotinib in combination with RORC inhibitors. The ALK+ cell lines K299, DEL and JB6 cells were left untreated, or treated with DMSO control, the RORC inhibitors SR2211 (7.5 μΜ), SR1903 (7.5 μΜ) and GSK805 (7.5 μM), or 25 nM crizotinib (Crizo) alone, or the different RORC inhibitors together with crizotinib. Cell death was analyzed by Annexin V-FITC/PI staining. The percentage of viable cells is shown. Experiments were performed in triplicates and results are shown as mean ± SD. One of three independent experiments is shown. *P < 0.05; ***P < 0.001; n.s., not significant
