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. 2017 Feb 8;7:40935. doi: 10.1038/srep40935

Figure 3. Ano5 gene knock-down with shRNA.

Figure 3

(A) Western blot of MC3T3-E1 cultures transfected with control or Ano5-specific shRNAs. Amounts of endogenous ANO5 protein increase over cell culture periods day 0 to day 21 in untransfected cell cultures (top row) and in cultures transfected with scrambled shRNA (middle row). Knock-down with Ano5-specific shRNAs is approximately 80% efficient (bottom row). (B) qPCR showing increasing Ocn, Cola1, Runx2 and osterix in knock-down MC3T3-E1 cultures compared to control groups. Relative expression to GAPDH. (C) Matrix and mineral formation in MC3T3-E1 cell cultures after ANO5 knock-down. Cells stained for alizarin red S. Mineral nodule formation is enhanced at days 14 and 21. Significant difference: *p < 0.05; **p < 0.01.