Figure 5
Characterization of SALL4 Degradation in hiPSCs by High Content Imaging. High content imaging analysis of SALL4 immunofluorescence with DAPI counterstaining in hiPSCs. (A,B) Time-course of SALL4 mean fluorescence intensity (MFI) in Gibco hiPSCs that were treated for the indicated time with 20 µM thalidomide (blue circles), 20 µM pomalidomide (red circles), 3 µM atRA (open circles), or DMSO (black circles) in mTeSR1 (A) or APEL2 + GSKi (B). (C) Dose-response characterization of thalidomide (blue circles), pomalidomide (red circles), lenalidomide (black circles), or atRA (open circles) on SALL4 MFI (normalized to the DMSO control) of Gibco hiPSCs that were differentiated in APEL2 + GSKi for 8 h in the presence of chemical. (D,E) Dose-response characterization of thalidomide (blue circles), pomalidomide (red circles), lenalidomide (black circles), or atRA (open circles) on SALL4 MFI of SALL4 mock (D) and SALL4G416A (E) XCL-1 hiPSCs that were differentiated in APEL2+ GSKi for 8 h in the presence of chemical. Data represent the mean +/− SEM from 2 independent experiments. Nonlinear regression analysis was performed using the 4-parameter [inhibitor] vs response variable slope plotting tool in GraphPad Prism.
