Extended Data Fig. 9: PICALM knockdown (KD) or knockout (KO) causes LD accumulation.

(a) Representative fluorescence staining of LD (BODIPY+) in iMG carrying risk or non-risk allele and in non-risk-CRISPRoff iMG. Scale bar, 50 μm. (b) to (e) Increased LD (BODIPY+) area, density per cell, size, and fluorescence intensity in iMG carrying the LOAD risk-allele (vs. non-risk) for both CD04 and CD09 lines. Note iMG with PICALM-CRISPRoff also show increased LD accumulation (vs. non-risk allele), mimicking the effect of the PICALM risk allele. (f) LD staining with LipidTox. Scale bar, 50 µm. (g) to (j) Increased LD density, size, area, and fluorescence intensity in iMG carrying the LOAD risk-allele (vs. non-risk). PICALM-CRISPRoff reduces LD density, size, area, and fluorescence intensity (vs. non-risk allele). For (b) to (e) and (g) to (j), each datapoint represents a single-well measurement (FOV) from one experiment; for each donor line (CD04 and CD09) and each condition (risk, non-risk, and CRISPRoff), data are from one experiment with 3 wells of differentiations each with 3-4 FOV. One-way ANOVA; Dunnett’s correction. (k) LD (BODIPY⁺) staining in PICALM-KO C20 cells and (l) quantification of LD number per cell. Scale bar, 25 µm. Each datapoint represents a single-well measurement (averaged from 5 FOV) from one experiment; data are from 3 experiments (n = 3). Two-sided unpaired Student’s t-test was used. For all comparisons, * P < 0.05, ** P < 0.01, ***, P < 0.001; mean ± s.e.m.