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. 2018 Apr 6;293(21):7942–7968. doi: 10.1074/jbc.RA118.002046

Figure 3.

Figure 3.

Conditional deletion of LRP6 in VSM increases NFATc4 mRNA, NFATc4 nuclear accumulation, and NFATc4 association with osteogenic genomic targets OPN and TNAP. A, primary aortic VSM cultures were prepared as detailed previously (6, 9) from LRP6-deficient SM22-Cre-LRP6(fl/fl);LDLR−/− mice and LRP6(fl/fl);LDLR−/− controls, and RNA was extracted for quantitation of NFATc messages as indicated. NFATc4 was significantly up-regulated with LRP6 deficiency. B, nuclear NFATc4 protein is also increased in primary aortic VSM cultures lacking LRP6. C, ChIP assays demonstrate that NFATc4 association with osteogenic OPN and TNAP promoters is increased with LRP6 deficiency. Associated with the TNF (tumor necrosis factor) control promoter was unaltered with LRP6 deficiency. D, RNAi targeting NFATc4 reduces osteogenic mRNA accumulation (OPN, TNAP, Msx2, Sox2, etc.) in cultures of aortic VSM cells. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001 versus corresponding control.