Extended Data Fig. 8: Exclusion of media lipid composition and apoptosis as confounding factors for observed lipidomic changes in MKs. Related to Fig. 4.
From: Critical shifts in lipid metabolism promote megakaryocyte differentiation and proplatelet formation
a, Distribution of main lipid classes from mature MKs (day 3, n = 3) as well as FBS (n = 11) used for cell culture supplementation. Lipid quantities were normalized to volume. b, Proteomic analysis of pro- and anti-apoptotic markers in MKs. A two-sided t-test was used for statistical analysis. Benjamini-Hochberg correction was applied to p-values using an FDR cut-off < 0.05 (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001). c, ATP cell vitality assay showing MKs treated with inhibitors. Ionomycin was used as positive control for apoptotic cells. The measured luminescence was normalized to the control for each day to display changes relative to the control baseline (dark grey). d, PS externalization during megakaryopoiesis. The two most abundant PS species were monitored. Externalization was calculated by dividing the amount of biotinylated PS on the cell surface by the total biotinylated PS, as described in the methods. Lipid quantities of day 1 were used as reference and set to 1. All other days were calculated as ratios relative to day 1 (n = 5). All data show the mean of at least 3 biological replicates. One biological replicate was comprised of 5 individual animals. In a-b, error bars represent standard deviations. In d, boxplot whiskers represent the minimum and maximum. The boundaries of the box represent the 25th and 75th percentile. Middle black lines represent the mean and black squares indicate the median.
