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. 2012 Jun;2(6):120063. doi: 10.1098/rsob.120063

Figure 3.

Figure 3.

RNAi depletion of USP11 inhibits TGFβ pathway signalling. (a) HEK293 cells were transiently transfected with siRNA targeting FoxO4 as control or USP11, starved for 4 h and stimulated with 50 pM TGFβ for 1 h prior to lysis. Extracts were resolved by SDS–PAGE and immunoblotted with antibodies against endogenous USP11, phospho-SMAD3 and SMAD3. (b) As in A except that HEK293 cells were transiently transfected with esiRNA targeting FoxO4 as control or USP11 and immunoblots against phospho-SMAD2 and SMAD2 were performed. Immunoblots are representative of two biological replicates each, using two sets of RNAi. (c) HEK293 cells were transiently transfected with siRNA targeting FoxO4 as control or USP11, starved overnight and stimulated for 4 h with 50 pM TGFβ. The expression of TGFβ-target genes PAI1 and GADD45B as well as USP11 knockdown were assessed by semiquantitative RT-PCR. Results are average of three biological replicates. Asterisk denotes statistical significance.