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. 2014 Aug 6;5:4619. doi: 10.1038/ncomms5619

Figure 7. Cdc42 expression regulates stiffening response and H3K9 me2 in melanoma cells.

Figure 7

(a) TRCs express lower level of Cdc42 compared with control melanoma cells and TRCs after culture on rigid plastic for 5 or 7 days. n=6; *P<0.05; two independent experiments. (b) TRCs regain cell stiffening responses after overexpression of Cdc42. Cells were plated on Collagen-I-coated PA gels with different stiffnesses or on glass-bottomed dishes. Experiments were carried out 6 h after plating. Relative changes in cell stiffness were represented with respect to the cell stiffness on 0.15 kPa PA gel as the baseline. TRC+empty vector: TRCs were transfected with pEGFP-N1 vector; TRC+Cdc42 overexpression: TRCs were transfected with Cdc42-EGFP plasmid. n>150 cells; *P<0.001. (c) Cdc42 knockdown inhibits cell stiffening in control cells and TRCs plated on plastic for 5 days. Relative changes in cell stiffness were represented with respect to the cell stiffness on 0.15 kPa PA gel as the baseline. n>100 cells; *P<0.001. Control: B16-F1 cultured on rigid plastic; TRC on plastic: TRCs were cultured on plastic for 5 days; Control+Cdc42 siRNA: control melanoma cells were transfected with Cdc42 siRNA; Control+Scrambled siRNA: control cells were transfected with scrambled siRNA; TRC on plastic+Cdc42 siRNA: TRCs were cultured on plastic for 5 days and then transfected with Cdc42 siRNA; TRC on plastic+Scrambled siRNA: TRCs were cultured on plastic for 5 days and then transfected with scrambled siRNA. Mean±s.e.m.; ‘*’ indicates significant difference compared with ‘0.15 kPa’. (d) Quantification analysis of western blotting of H3K9me2 and me3 after Cdc42 knockdown in control melanoma cells. Mean±s.e.m.; n=3 independent experiments; *P<0.05.