Fig 2: Western blot analysis of expression of core targets of DMAG. The expressions of ITGA2B, ITGB3, VWF, PLEK and p-Akt were detected by western blot after HEL cells were treated with DMAG (10, 20 and 40 µM) for 4 days. The data represent the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, compared to the control group

Fig 3: DMAG induces the differentiation of HEL cells. a The expression of CD41 and CD61 was detected by flow cytometry after the HEL cells were treated with DMAG (10, 20 and 40 µM) for 6 days. The histogram shows the percentage of CD41+CD61+ cells in each group. The data represent the mean ± SD of three independent experiments; ***p < 0.001, compared to the control group. b The expression of CD41 and CD42b was measured by flow cytometry after the HEL cells were treated with DMAG (10, 20 and 40 µM) for 6 days. The histogram shows the percentage of CD41+CD42b+ cells in each group. The data represent the mean ± SD of three independent experiments; **p < 0.01, ***p < 0.001, compared to the control group. c Flow cytometry analysis of DNA ploidy after the cells treated with DMAG (10, 20 and 40 µM) for 6 days. The histogram shows the percentage of DNA ploidy in each group. The data represent the mean ± SD of three independent experiments; **p < 0.01, ***p < 0.001, compared to the control group. d HCS analysis of DNA ploidy after the cells treated with or without DMAG (10, 20 and 40 µM) for 6 days. Cells were marked with blue, turquoise, green, orange and red. The different colors represent different phases of the cell cycle. Blues indicate G0/G1 (2 N) cells. Turquoises indicate S (2 N) cells. Greens represent G2 (= 4 N) cell. Oranges represent early M (= 4 N) cells. Reds represent late M (2 N) cells. The histogram shows the percentage of DNA ploidy in each group. The data represent the mean ± SD of three independent experiments; ***p < 0.001, compared to the control group

Fig 4: DMAG administration enhances recovery of platelet level and function in mice with thrombocytopenia. a The KM mice were irradiated by X-ray (4 Gy) and then treated with normal saline, TPO (3000 U/kg), or DMAG (5 mg/kg) for 14 days, respectively. The number of platelets were counted on day 0, 3, 7, 10, and 14. Each group contains 12 mice (6 male mice and 6 female mice). The data represent the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, compared to the corresponding model group. b MPV was measured on day 0, 3, 7, 10, and 14 in each group. c Representative images of H&E staining of BM from normal, model, TPO and DMAG groups. The yellow circles indicate megakaryocytes. Scale bar: 100 µm. The histogram indicates the number of megakaryocytes in each group. The data represent the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01, compared to the model group. d Immunohistochemical analysis of the expression of VWF in BM cells of each group. Blue represents cell nucleus. Brownness represents the expression of VWF. The histogram indicates the number of megakaryocytes in each group. The data represent the mean ± SD of three independent experiments; **p < 0.01, ***p < 0.001, compared to the model group. e The expression of CD41 and CD61 was detected by flow cytometry in BM cells of each group. The histogram indicates the percentage of CD41+CD61+ cells in each group. The data represent the mean ± SD of three independent experiments; ***p < 0.001, compared to the model group. f Tail bleeding time was measured after the distal tail was cut in each group. The data represent the mean ± SD of three independent experiments; **p < 0.01, compared to the model group. g Under the stimulation of ADP, platelet aggregation was measured by a turbidimetric aggregation-monitoring device. The histogram shows the maximum aggregation amplitude of platelets in each group. The data represent the mean ± SD of three independent experiments; **p < 0.01, compared to the model group. h Micrographs of collagen-coated slides with the same number of platelets perfused. The red represents platelets. The histogram shows the average coverage of red fluorescence on the whole surface by ImageJ software. The data represent the mean ± SD of three independent experiments; ***p < 0.001, compared to the model group