Figure 3.

Relationship of CAIX to NF-κB. (A) CAIX depletion reduces NF-κB activity. Luciferase assay monitoring the activity of the NF-κB reporter vector in hypoxic cells with induced CAIX expression (100%) in comparison to CAIX-knockout MCF7-CA9-KO, MDA-MB-321-CA9-KO, and transiently silenced MCF7-siCA9 cells. (B) Impact of CAIX enzymatic inhibition by HSFA (100 µM) on CA9, LIN28B, PDK1, and NF-κB mRNA expression in hypoxic MCF7 cells. (C) Effect of modified acidic medium pH 6.7 on the set of the same genes mRNA expression in hypoxic MDA-MB-231 and MCF7 cells. mRNA expression was normalized to actin and presented as a percentage of expression in the control medium. The results represent the mean of three independent biological experiments performed in triplicates. p > 0.05 was considered nonsignificant (ns), p < 0.05 is denoted as *, p < 0.01 as ** and p < 0.001 as ***.