Fig. 4

The role of gene expression in 4SC-202-induced arrest and cytotoxicity. a, b To evaluate the extend of gene expression changes induced by FK228 and different concentrations of 4SC-202, we applied NanoString nCounter® analysis with mRNA derived from the indicated CTCL cell lines after 24-h treatment with 4SC-202 and FK228. A NanoString Panel allowing quantification of mRNA from 519 different kinases and 6 housekeeping genes was used. Following normalization, changes relative to DMSO treated control cells were calculated and are displayed in a with the indicated color code. Very low expressed genes (< 0.03% of GAPDH) very excluded from further analysis and are displayed as not evaluable. b The percentage of genes that were more than twofold up- or downregulated are depicted. c The indicated cell lines were incubated for 72 h with 4SC-202 or FK228. Cellular DNA was stained by propidium iodide and the increase in sub-G1 cells relative to control cells is depicted. d To evaluate the necessity of de novo gene expression for the 4SC-202-induced G2/M arrest, HTB-176 cells were first arrested in prometaphase by a 12-h treatment with 100 nM nocodazole (Noc). Then nocodazole was removed and cell cycle progression in the presence or absence of the transcription inhibitor Actinomycin D (Act.D) and/or 4SC-202 was assessed by propidium iodide staining