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. 2021 Oct 12;10(10):2721. doi: 10.3390/cells10102721

Figure 5.

Figure 5

The TIMP-1–CD63 axis regulates CAIX expression and extracellular acidosis in aggressive breast cancer cells. (A) RT-PCR and (B) immunoblot analyses of CAIX and CAXII expression in control (shCONT), shTIMP-1, and shCD63 MCF10CA1h cells. GAPDH and β-actin were analyzed as controls for RT-PCR and immunoblot analysis, respectively. (C) Changes in the extracellular pH in control (shCONT) and shTIMP-1 or shCD63 MCF10CA1h cells. Change in extracellular pH was normalized to cell number to account for cell proliferation. Bars represent the mean ± SD. * and ** p < 0.05 comparing shCONT to shTIMP-1 and shCONT to shCD63, respectively. (D) Extracellular acidosis was monitored using the pH sensitive DF fluorescein dye. Parental MCF10A cells were loaded with the DF fluorescein dye followed by incubation with the conditioned media from control (shCONT), shTIMP-1, or shCD63 MCF10CA1h cells. Scale bar represents 40 µm.