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. 2000 Feb 25;97(6):2791–2796. doi: 10.1073/pnas.050574297

Figure 4.

Figure 4

RT-PCR experiment to analyze IL-6, IL-8, and IL-15 expression in wnt5a-transfected (transient) normal synovial fibroblasts and determination of transfection efficiency. (A) Lane 1, DNA standard PhiX174; lanes 2–5, IL-6-, IL-8-, IL-15-, and G3PDH-specific RT-PCR products from wnt5a-pcDNA3-transfected normal synovial fibroblasts; lanes 6–9, IL-6-, IL-8-, IL-15-, and G3PDH- specific RT-PCR products from empty vector transfected normal synovial fibroblasts; lanes 10–13, IL-6-, IL-8-, IL-15-, and G3PDH-specific RT-PCR products from untransfected normal synovial fibroblasts. (B) Bar graph showing fold increase in IL-6, IL-8, and IL-15 gene expression (measured by PCR product intensity) on transfection of normal synovial fibroblasts with wnt5a expression construct. Fold increase with wnt5a is compared with the effects produced by transfection by the empty vector. This represents an average of five different experiments. For each experiment, the same amounts of RNA were used from the wnt5a- and empty vector-transfected cells. G3PDH expression was used an internal control. (C) Western blot of lysate from wnt5a-HA-transfected synovial fibroblasts. Lane 1, ≈60 kDa wnt5a-HA protein from wnt5a-HA-transfected synovial fibroblasts; lanes 2 and 3, no band observed from untransfected synovial fibroblasts.