Extended Data Fig. 2: Fatty acid uptake and synthesis affected MK differentiation but not mitochondria metabolism. | Nature Cardiovascular Research

Extended Data Fig. 2: Fatty acid uptake and synthesis affected MK differentiation but not mitochondria metabolism.

From: Efficient megakaryopoiesis and platelet production require phospholipid remodeling and PUFA uptake through CD36

Extended Data Fig. 2: Fatty acid uptake and synthesis affected MK differentiation but not mitochondria metabolism.The alternative text for this image may have been generated using AI.

(a) Fetal liver-derived HSPCs were cultured with TPO and treated with the ACSL inhibitor Triacsin C at indicated doses. CD41+ and CD41/CD42d+ cells were quantified using flow cytometry, n = 5, one-way ANOVA – Dunnett’s test. Data are presented as mean +/− SD. (b) Cytotoxicity assay was performed on day 4 after fetal liver MKs were treated with Triacsin C. Control=cells treated with the vehicle; positive control=cells lysed with TritonX-100. n = 4. Data are presented as mean +/− SEM (c) Fetal liver derived HSPCs were cultured with TPO and treated with the ACC inhibitor PF-05175175 at indicated doses. CD41+ and CD41/CD42d+ cells were quantified using flow cytometry, n = 5, one-way ANOVA – Dunnett’s test. Data are presented as mean +/− SD (d) Cytotoxicity assay was performed day 4 after fetal liver MKs treated with PF-05175175. Control=cells treated with the vehicle; positive control=cells lysed with TritonX-100. n = 4. Data are presented as mean +/− SEM (e) Fetal liver derived HSPCs were cultured with TPO and treated with the FASN inhibitor Cerulenin at indicated doses CD41+ and CD41/CD42d+ cells were quantified using flow cytometry, n = 5, one-way ANOVA – Dunnett’s test. Data are presented as mean +/− SD (f) Cytotoxicity assay was performed day 4 after fetal liver MKs were treated with Cerulenin. Control=cells treated with the vehicle; positive control=cells lysed with TritonX-100, n = 4. Data are presented as mean +/− SEM (g) Representative graph of mitostress assay. MKs were treated with Triacsin C (T4540, Sigma), PF-05175175 (PZ0299, Sigma), Cerulenin (C2389, Sigma) at the indicated concentrations in complete media for 90 min prior the measurement. Oxygen consumption rates were measured in accordance with manufacturer instructions (Agilent/Seahorse Bioscience) n = 5 biological replicates (h) Individual parameters for basal respiration, maximal respiration, and spare respiratory capacity were measured and analyzed. n = 5 biological replicates, one-way ANOVA. Data are presented as mean +/− SD.

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