FIGURE 6.
Lentiviral knockdown of HAS3 and HAS2 cause FAK cleavage and inhibit proliferation and migration. A, lentiviral knockdown of HAS3 and HAS2 was performed as described in Fig. 5. Increased cleavage of tFAK resulting in the loss of total intact 125-kDa FAK occurred after shHAS3 expression. This effect was present but less pronounced after knockdown of shHAS2. Quantitative analysis of 125-kDa tFAK and degraded tFAK (<125 kDa) after normalization to tubulin and to scrambled (scr) controls (n = 3, mean ± S.E.). *, p < 0.05 versus scrambled shRNA. B, representative immunoblots of tAKT, pAKT, tERK, and pERK and quantitative data after densitometric scanning and normalization to tubulin and to scrambled controls (n = 3, mean ± S.E.). *, p < 0.05 versus scrambled shRNA. The effects of HAS knockdown were observed 5 days after infection and compared with a nontargeting lentivirus as control. C, effect of shHAS3 and shHAS2 on migration toward FCS as determined in a 24-well microchemotaxis assay and on DNA synthesis as determined by [3H]thymidine incorporation in response to FCS expressed as cpm per total cellular protein (n = 3, mean ± S.E.). *, p < 0.05, ***, p < 0.001 versus scrambled shRNA. scr, scrambled.