One part of my experiment was to analyse the phosphorylation status of Dvl2. I had transfected cell lines with Wnt6 and Dvl2 to see the effect wnt 6 would have on Dvl2. Following transfection protocols cells were starved for 24 hours and then harvested. I confirmed successful transfection of samples with Dvl2 and Wnt6 using western blot analysis. The conditions were as follows:
(Control) (Dvl2) (Wnt6) (Dvl2 and Wnt6 co-transfection)
One problem is that Dvl2 expression in the conditions were not equal even though B-actin showed loading to be equal.
The expression of Dvl2 in the (Dvl2 + Wnt 6) were lower than in (Dvl2 alone). I don't understand this, does this mean that regardless of protein loading being equal Wnt 6 is downregulating expression of Dvl2 or that transfections worked better in (Wnt6) condition than in (Dvl2+Wnt 6).
I need help in analyzing these results. What do they mean?