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. 2016 Dec 30;8(6):10225–10237. doi: 10.18632/oncotarget.14379

Figure 4. Generation of CA9-ko cells using CRISPR-cas9 to create single and NHE1/CA9 double knockout cell lines: impact on proliferation in Nx and Hx.

Figure 4

A. WB showing protein expression patterns for HIF1, NHE1, CA9, CA12 and loading controls (Actin, ARD1) in CA9-ko, NHE1-ko and NHE1/CA9-dko generated cells in comparison to WT cells from Nx and Hx growth conditions. B. Cellular proliferation assays from day 6 represented as fold increase standardized to the starting number of cells at 24h post-plating. Proliferation in Nx and Hx are presented on the same figure. The general CA inhibitor acetazolamide (ACTZ, 100μM) was used for comparative purposes. Statistical differences are indicated by an asterisk ‘*’ as compared to WT cells with analyses occurring in Nx and Hx assays individually. P values less than 0.05 are represented as ‘*’ while p values less than 0.001 are represented by ‘**’. Proliferation data were obtained from duplicate measurements on 3-4 independent experiments.