Fig 2: Selectivity and efficiency of JAK-targeting siRNAs. Selectivity of the sorted siRNAs was assessed on PC3-JAK3-GFP-engineered cells transfected using lipofectamine RNAiMAX and different siRNAs at 10 nM. Representative western blot analysis is shown on the top panel, with the corresponding quantification at the bottom [N = 4 independent experiments] for the expression of JAK1 [A], JAK3 [B], JAK2 [C] or TYK2 [D] proteins. For A, * represents respectively p-values = 0.0307 and 0.0324; for B, * represents respectively p-values = 0.0477 and 0.0456; for D, * represents respectively p-values = 0.0212 and 0.0359. [E] Representative western blot of PC3 cells transfected by JAK1-targeting siRNAs at different concentrations ranging from 0.5 to 12.5 pM. [F] Quantification of JAK1 protein expression [N = 3 independent experiments] from the experiment shown in E. Quantification by RTqPCR of JAK1 mRNA levels in PC3 cells [G, H] or JAK3 mRNA levels in PC3-JAK3-GFP-engineered cells [I, J] transfected using lipofectamine RNAiMAX and siRNAs at concentration ranging from 0.5 to 12.5 pM [N = 3 independent experiments]. GAPDH was used as a housekeeping gene.