Fig. 8

Inhibiting calcification in 4T1 cells with acetazolamide. The 4T1 cells were incubated with osteogenic cocktail (OC) supplemented with 1 mM acetazolamide (AZ). a The 4T1 cells cultured with OC supplemented with DMSO in the presence or absence of AZ were stained with AR-S and were photographed at the original magnification and at ×100 magnification. b The 4T1 cells, cultured with OC supplemented with DMSO in the presence or absence of AZ, were stained with cetylpyridinium chloride and were quantified by measuring their absorbance at 562 nm. c CA1 transcription in 4T1 cells was detected by real-time PCR. d CA1 protein expression in 4T1 cells was detected using western blot analysis. e The protein expression level was normalised to the expression of GAPDH. * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001