Fig. 3

CITK scaffold activity, but not kinase activity, is required for regulation of BRCA1 levels during neurodevelopment.

a Schematic representation of the protocol used to obtain neural progenitor cells and forebrain organoid from pluripotent stem cells. Representative images of proliferating neural progenitor cells (b) or rosettes from DD35 organoid sections (e), carrying wild type, frameshift, or kinase inactive variants of CIT gene, immunostained for BRCA1 and counterstained with DAPI. Scale bars: 10 μm and 50 μm, respectively. c, d, f, g Quantification of BRCA1 foci per nucleus in each genotype and condition. h Western blot analysis of total lysate obtained from P4 cerebellum and P4 cortex of mice carrying wild type, frameshift, or kinase inactive variant of Cit gene. The levels of CITK and BRCA1 were analyzed, and the internal loading control was α-tubulin (α-TUB). i, j Quantification of BRCA1 levels in each genotype. Each dot indicates an independent biological replicate. Error bars, SEM. *P < 0.05, ***P < 0.001; unpaired two-tailed Student’s t-test for western blots, Mann–Whitney U test for BRCA1 foci. A.U. arbitrary unit.