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. 2019 Sep 27;9:992. doi: 10.3389/fonc.2019.00992

Figure 5.

Figure 5

Silencing of RhoB GTPase does not affect the migration of brain cancer cells under mechanical compression. (A) H4 and A172 brain cancer cells were transiently transfected with siRNA against RhoB and were compressed by 4.0 mmHg in 2% FBS containing DMEM. Total RNA was then isolated and RhoB mRNA expression was quantified by qPCR. Each bar indicates the mean fold change ±SE of three independent experiments (number of total replicates n = 9). Asterisk (*) indicates a statistically significant difference (p < 0.05). (B) Western Blotting showing that RhoB protein expression has been successfully reduced in compressed siRhoB-treated cells (lanes 2 and 4) compared to compressed control cells (lanes 1 and 4). Protein expression was quantified using ImageJ software and the fold change is indicated in gray font. (C) H4 and A172 cells knockdown for siRhoB were compressed by 4.0 mmHg in low-serum medium and then subjected to a scratch wound healing assay for 16 h. Control cells were treated with non-specific control siRNA (NSC). Scale bar: 0.2 mm. (D) Graph showing the percentage wound closure as quantified using ImageJ software (≥3 different fields/condition; ≥3 independent experiments number of total replicates n ≥ 10). No statistically significant differences in wound closure were observed between both H4 and A172 siRhoB-treated cells and the respective NSC-treated cells.