Figure 3.
AGK regulates thrombocytopoiesis through a nonmitochondrial function. (A) Genotyping of AGK-G126E PM mice by sequencing. (B) The levels of AGK in the platelets of AGK-G126E PM mice were tested by western blot analysis. (C) Platelet, RBC, and WBC counts in WT and AGK-G126E PM mice (n = 4). (D) Immunofluorescence images of α-tubulin in PPF-megakaryocytes derived from the fetal livers of WT mice and AGK-G126E PM mice. The scale bars represent 50 μm. Statistics for the PPF-megakaryocytes/total megakaryocytes derived from the fetal livers of WT mice and AGK-G126E PM mice are shown (n = 10). (E) Extracellular flux analysis of the OCRs of the platelets of WT and PM mice; 2 μM oligomycin, 0.25 μM FCCP, and 1 μM rotenone/antimycin A were added at the indicated time points. The OCR was normalized to the protein amount (n = 6). (F) Extracellular flux analysis of the OCRs of the platelets of Agkf/f and Agk−/− mice; 2 μM oligomycin, 0.25 μM FCCP, and 1 μM rotenone/antimycin A were added at the indicated time points. The OCR was normalized to the protein amount (n = 4).

AGK regulates thrombocytopoiesis through a nonmitochondrial function. (A) Genotyping of AGK-G126E PM mice by sequencing. (B) The levels of AGK in the platelets of AGK-G126E PM mice were tested by western blot analysis. (C) Platelet, RBC, and WBC counts in WT and AGK-G126E PM mice (n = 4). (D) Immunofluorescence images of α-tubulin in PPF-megakaryocytes derived from the fetal livers of WT mice and AGK-G126E PM mice. The scale bars represent 50 μm. Statistics for the PPF-megakaryocytes/total megakaryocytes derived from the fetal livers of WT mice and AGK-G126E PM mice are shown (n = 10). (E) Extracellular flux analysis of the OCRs of the platelets of WT and PM mice; 2 μM oligomycin, 0.25 μM FCCP, and 1 μM rotenone/antimycin A were added at the indicated time points. The OCR was normalized to the protein amount (n = 6). (F) Extracellular flux analysis of the OCRs of the platelets of Agkf/f and Agk−/− mice; 2 μM oligomycin, 0.25 μM FCCP, and 1 μM rotenone/antimycin A were added at the indicated time points. The OCR was normalized to the protein amount (n = 4).

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