Restoration of E-cadherin expression in NMuMG cells is dependent on levels of ZEB1 and ZEB2. NMuMG cells were incubated for 48 hours with 100 pM TGF-β1 to induce EMT. Thereafter, the cells were incubated for an additional 24 hours with the indicated inhibitors: (lane 3), 1 μM SB431542; (lane 4), 1 μM Y27632; (lane 5), 1 μM SB431542 and 10 μM SP600125; and (lane 6), 1 μM SB431542 and 1 μM Y27632. NMuMG cells were also infected with shRNAmir-targeting ZEB1 and ZEB2 lentiviruses followed by incubation for an additional 24 hours without TGF-β1 (lane 7), with 100 pM TGF-β1 (lane 8), or with 100 pM TGF-β1 plus 1 μM Y27632 (lane 9). Levels of ZEB1, ZEB2, E-cadherin, and β-actin protein in the cells were assessed by immunoblot analysis. Dash indicates protein present at level below background; plus sign indicates protein present at clearly detectable level. Similar findings were observed in two other independent experiments.