Human G795A and G795C iPSC-derived microglia are resistant to multiple CSF1Ri. (A) Chromatograms of CRISPR-modified isogenic set of CSF1R-G795A, G795C, and G795V human iPSC lines. (B and C) Quantification of iMG cell death by fluorescent Caspase 3/7 Dye for Apoptosis over 24 h in culture with complete medium treated with 0.1% DMSO, 250 nM, 500 nM, or 1 µM Edicotinib (B) and equivalent concentrations of BLZ945 (C). (D) Live (Calcein-AM) versus dead (ethidium homodimer-1) percent quantification of WT and G795A microglia cultured with 0.1% DMSO or 500 nM of PLX3397, PLX5622, Edicotinib, or BLZ945. Data captured from four images/well from n = 6 independent wells using an Incucyte S3 live-cell imaging system. Data represented as percent mean values. (E–G) Percent confluency of iMG normalized to t = 0 h over 24 h in culture with complete medium treated with 0.1% DMSO or 500 nM PLX3397 (E), PLX5622 (F), Edicontinib (G), or BLZ945 (H). Data captured from four images/well from n = 6 independent wells using an Incucyte S3 live-cell imaging system. Data represented as mean values ± SEM. (I and J) Hierarchical clustering and Pearson correlation of normalized RNA counts of the whole transcriptome (I) and a 249-gene microglia signature (J) between WT (black), G795A (green), and G795C (blue) iMG cultured with 0.1% DMSO or 250 nM PLX5622 (n = 4 replicates per cell line, per condition). Lightest shade of blue in I and J indicates correlations of R = 0.93; darkest shade of blue indicates R = 1.0 correlation.