Figure 2.
ShRNA transfection. (A) GFP-expressing control shRNA demonstrates shRNA stable transfection efficiency; Bar = 25 μm. (B) Western blots were performed for control, GPC1, and SDC1 shRNA transfections. We assessed whole cell protein levels of GPC1 and SDC1. β-actin was probed as a loading control. In the left panel of Western blots lane 1 is an untreated control, lane 2 is a scrambled shRNA control, and lane 3 is the GFP-expressing shRNA control; all bands are similar for all conditions, as expected. In the middle panel of Western blots lane 4 is an untreated control and lane 5 is the GPC1 shRNA treatment; the change in GPC1 is expected and it is accompanied by a change in SDC1. In the right panel of Western blots lane 6 is an untreated control and lane 7 is the SDC1 shRNA treatment; the change in SDC1 is expected and there is no change in GPC1. (C, D) Plots are based on GPC1 shRNA and SDC1 shRNA knockdown of whole cell GPC1 and SDC1 protein as determined using Western blots and densitometric analysis. **P ≤ 0.01 or ***P ≤ 0.001; n = 6.