Figure 8

Knockdown of COL3A1 suppresses the proliferation of neurofibromin-deficient cells. (a) Phase-contrast microscopy of sNF96.2 cells that had been transfected with control (GAPD) or COL3A1 siRNAs for 2 days. Scale bar, 100 µm. (b) Phase-contrast microscopy of neurofibroma cells or DFAT cells from NF1 patients 1 and 2 that had been transfected as in a. Scale bar, 100 µm. (c) Quantitative RT-PCR analysis of COL3A1 and SOX2 expression in neurofibroma cells of patient 1 that had been exposed to tranilast (250 µM) for 20 days. Data are means ± s.d. for triplicates from a representative experiment. ***P < 0.001 versus corresponding control value (Student’s unpaired t test). (d) Tranilast-resistant neurofibroma cells derived from patient 1 were transfected with control (GAPD) or COL3A1 siRNAs for 1 day and then exposed to tranilast (250 µM) or DMSO vehicle for 48 h, after which the cells were examined by phase-contrast microscopy. Scale bar, 100 µm. The number of viable cells and the percentage of viable cells were also measured on the basis of trypan blue exclusion. Data are means ± s.d. for triplicates from a representative experiment. *P < 0.05, **P < 0.01 (Student’s unpaired t test); ns, not significant.