Fig. 6

Phenotypic rescue of SETBP1 deficiency by pharmacological interrogation of Wnt signaling. A Experimental scheme. Differentiation cultures under basal condition (control) or exposed to 2uM XAV939 from day 11 to day 21. B Western blot analysis at day 35 for the effects of XAV treatment on WNT signaling proteins. Data was obtained from 2 independent differentiations analysed in duplicates or triplicates and shown as relative levels to the WT. Student’s T test was used to compare the expression between the two lines, B-catenin basal P = 0.033, XAV P = 0.542, S552 basal P = 0.023, XAV P = 0.906, S674 basal P = 0.024, XAV P = 0.554, LRP6 basal P = 0.153, XAV P = 0.644, P-LRP6 basal P = 0.004, XAV P = 0.012. C Effect of XAV treatment on cell cycle profile at day 35. Data presented as mean ± s.e.m of 2 independent experiments in triplicates. Student’s T test, one tail, was used to compare the expression between the two lines and the two conditions (WT basal vs. XAV G0-G1 P = 0.029, S P ≥ 0.05, G2-M P ≥ 0.05, SETBP1-/- basal vs. XAV G0-G1 P = 0.021, S P ≥ 0.05, G2-M P = 0.0045. D Immunofluorescence images of cultures in basal (DMSO) or XAV treated conditions at day 20 and 30. Cell nuclei were labelled by DAPI. Scale bar: 100uM. Bar- graphs showing quantification of FOXG1 (green) positive NPS at day 20 and CTIP2 (green), TBR1 (red) and NeuN (green) positive neurons at day 30. Student’s T test was used to compare the expression between the two lines, FOXG1⁺ cells: basal P = 0.002, XAV P = 0.001; CTIP2⁺ cells: basal P = 0.029, XAV P = 0.214; TBR1⁺ cells: basal P = 0.672 XAV P = 0.258, NeuN⁺ cells: basal P = 0.038, XAV P = 0.333 (*p ≤ 0.05, **p ≤ 0.01). E Schematic illustration depicting the role of SETBP1 in regulating NPC pool expansion and forebrain fate induction