Fig. 5: VAC14 is required for human erythroid differentiation.

A Mean sgRNA abundance in library, HUDEP-2 cells prior to differentiation, and HUDEP-2 cells at day 12 of differentiation, obtained from the secondary custom CRISPR screen. B Fold change in counts of HUDEP-2 cells grown in maintenance media following transduction with one of 3 shRNAs targeting VAC14 (resulting in 50–70% reduction in VAC14 mRNA levels) or scramble control shRNA. Statistical analysis was performed compared to scramble control, using two-way ANOVA followed by Dunnett’s multiple comparison test (n = 3 biological replicates per condition, data represented as mean ± standard deviation). C Fold change in counts of erythroid cells differentiated from CD34+ HSPCs following transduction with one of 3 shRNAs targeting VAC14 or scramble control shRNA. Statistical analysis was performed using two-way ANOVA followed by Dunnett’s multiple comparison test (n = 3 biological replicates per condition, data represented as mean ± standard deviation). D Erythroid differentiation as assessed by flow cytometry (E) Cytospin images of day 18 erythroid cells differentiated from HSPCs following transduction with one of 3 shRNAs targeting VAC14 or scramble control shRNA. Images are representative of 3 independent experiments. ***p < 0.001; ****p < 0.0001. Source data are provided in Source Data file.