Fig. 6

Lovastatin inhibition of HMGCR prevents cells adaptation to shear stress environment. Erythroblasts (EBL) of three different donors (n = 3) were incubated with different concentrations of lovastatin on day 0 of differentiation and cultured in static and dynamic conditions. Erythroid cells were harvested on day 12 of differentiation and analysed by flow cytometry. A) Representative example of living gate strategy plotting forward scatter (FSC x-axis) vs side scatter (SSC y-axis) in cells differentiated in static and dynamic conditions treated with different concentrations of lovastatin. B) Maturation states of cells differentiated in static and dynamic conditions incubated with different concentrations of lovastatin according to antiCD49d-PB (BD Biosciences, San Jose, CA, US) (Alexa Fluor 405, y-axis) and antiCD235a-PE (OriGene Technologies, Rockville, MD, USA) (x-axis), staining. C) Quantified percentage of cells in the living gate, averaged for 3 donors (n = 3). Gating strategy to identify the living cells gate is described in Supplemental Fig. 4). D) Quantified maturation states of cells differentiate in static and dynamic incubated with different concentrations of lovastatin were measured as described in Fig. 1A. Percentage of cells in gates was averaged for 3 donors (n = 3). ND indicates not-detectable measurement due to the low percentage of cells in the living gate. E) Representative picture of pelleted cells on day 12 of differentiation incubated with different concentration of lovastatin. C and D) 2way Anova test was performed, with *** p < 0.001 ** p < 0.01 and *p < 0.05. Unless marked, no significance was observed.