Figure 4: Effect of PPARγ knockdown on the growth characteristics of HTh74 cells. (a) Cell growth rate was assessed by counting viable cells transduced with scrambled (scr) or PPARγ shRNA at 2-day intervals. Data is represented as fold cell number over day 0 (6 expts. ± SEM, *, , 2-way ANOVA calculated over the entire growth curve). (b) scr-(white bars) and PPARγ shRNA-(black bars) transduced HTh74 cells were growth arrested in 0.5% FBS for 24 h and analyzed by flow cytometry 24 h following subsequent growth in 5% FBS. Flow data (3 expts.) was quantitated for % of cells in G1, S, and G2/M phases as shown (*, G1- ; S, ; G2/M, : paired -test). (c) 50 μg of total cellular protein from HTh74 cells transduced with either scrambled or PPARγ-specific shRNA was separated on a 10% SDS-PAGE gel, transferred to PVD, and probed with antibodies against the corresponding cell cycle protein whose source is described in Section 2.4 followed by the appropriate secondary antibody. β-actin was quantitated as a loading control.