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. 2011 Nov 4;6(11):e27155. doi: 10.1371/journal.pone.0027155

Figure 1. Efficiency and specificity of Ac45 siRNA knockdown in osteoclasts.

Figure 1

(A) Five Ac45 siRNA oligonucleotides were tested for their ability to silence Ac45 mRNA expression. Semi-quantitative RT-PCR for Ac45 gene expression was carried out 48hrs post-transfection of siRNA oligos (100nM) into pre-osteoclasts. Expression levels were normalized to 18S and expressed as percentage of control ± SEM (n = 3). (B) Specificity and efficiency of Ac45 silencing. mRNA from pre-osteoclasts transfected with Ac45 siRNA for 48hrs was subjected to semi-quantitative RT-PCR analysis using specific primers for V-ATPase V0 domain subunits a3 and d2, CTR and 18S. (C) Quantitative analysis of gene expression of Ac45 and V-ATPase V1 (B2 and E1) and V0 (a3, d2, c″ and c) subunits following Ac45 silencing using real-time qPCR. Data represented as relative mRNA level normalized to 36B4 control ± SEM (n = 3). (D) Ac45 protein level after Ac45 silencing in pre-osteoclasts for 48hrs. *P-values<0.05.