Figure 4.
The quiescent cancer cells are characterized with intracellular acidification. (A) Cells were treated with PTX for 7 days then partially released to reactivation (left), the reactivated cells were treated with higher dose of PTX for 3 days (right), intracellular pH (pHi) was examined by BCECF-AM assays kit according to the manual protocol. Data are shown as mean ± SD of three independent experiments, * P<0.05, (B) NHE1 proteins in these cells were detected by Western blot. (C-E) The reactivated cells were treated with 100 µM Cariporide for 24h, (C) the intracellular pH was examined by BCECF-AM assays kit, data are shown as mean ± SD of three independent experiments, * P<0.05, (D)The expression of Cyclin D1 were detected by Western blot, GAPDH was used as loading control, (E) Cell proliferation was detected by EdU incorporation assay, data are shown as mean ± SD of three independent experiments, * P<0.05.(F) The cell growth curve during 3 days culture.