Fig. 2
PP2A-Tws dephosphorylates Map205 at Ser283. (A) Map205 is hyperphosphorylated at Ser283 after depletion of RNAi depletion of Tws. Cells expressing Myc-Map205254 − 400 WT or S283A were transfected with dsRNA against Tws or Non-Target (NT). After 4 days, cells were analyzed by Western blotting using Phos-tag as indicated. Left: results from a representative experiment. Right: Normalized ratios of phosphorylated/unphosphorylated Myc-Map205254 − 400 band intensities. Averages of 3 experiments are shown. All error bars: S.D. *** p < 0.001, **** p < 0.0001 from paired t-tests. (B) PP2A-Tws dephosphorylates a Map205 peptide at Ser283 in vitro. Left: Flag-Tws or Flag-GFP were immunoprecipitated from cells and products were analyzed by Western blotting. Right: Dephosphorylation of the pSer283 Map205 peptide by Flag-Tws as a function of time (violet curve). Values obtained with Flag-GFP were subtracted. The Flag-Tws associated phosphatase activity is largely abrogated by the addition of LB100, a PP2A inhibitor (orange curve). Experiment repeated 3 times. Averages of technical triplicates from a representative experiment are shown. AU: arbitrary units. **** p < 0.0001 from unpaired t-tests