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. 2015 Feb;185(2):485–495. doi: 10.1016/j.ajpath.2014.09.021

Figure 6.

Figure 6

Stable silencing of Sp1 or Sp3 using lentivirally delivered shRNAs decreases xylosyltransferase-1 (XT-1) expression in human nucleus pulposus (NP) cells. A and B: Real-time PCR analysis of human NP cells transfected with LV-sh-control (LV-shC) or LV-shSp1 (A) or LV-shSp3#2 (B). XT-1 mRNA expression was significantly suppressed by LV-shSp1 and LV-shSp3#2 compared with cells transduced with control lentivirus (LV-shC). Strong suppression in Sp1 and Sp3 mRNA expression was observed with corresponding gene-targeting shRNAs. C–F: Western blot (C and E) and corresponding densitometric (D and F) analyses of NP cells transduced with LV-shC or LV-shSp1 (C and D) or LV-shSp3 (E and F). XT-1 expression levels were suppressed by LV-shSp1 and LV-shSp3#2 compared with cells transduced with control shRNA. Targeting sequence shSp3#1 did not result in the suppression of Sp3 levels. As expected, transduction of cells with shSp1 or shSp3#2 showed a corresponding decrease in Sp1 or Sp3 protein levels. Data represent means ± SEM of three independent experiments performed in triplicate. ∗P < 0.05.