Fig. 8
Oxymatrine induces megakaryocyte (MK) differentiation by activating stimulator of interferon genes (STING)/nuclear factor-kappaB (NF-?B) pathway. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for the potential pathway by which oxymatrine promotes MK differentiation. (B?E) Western blot (WB) analysis of the expression of pathway: extracellular regulated kinase 1/2 (ERK1/2)/STING/inhibitory subunit of nuclear factor kappa B alpha (I?B?)/NF-?B by which oxymatrine promotes MK differentiation. Representative immunoblot images and biochemical quantification of the ERK1/2 (B), STING (C), I?B? (D), and NF-?B (E) pathway after the treatment with oxymatrine (10, 20, and 40 ?M) in Meg-01 cells for five days (n = 3 per group). (F?I) Representative immunoblot images and biochemical quantification of the transcription factors (TFs) associated with MK differentiation: runt-related transcription factor 1 (RUNX1) (F), nuclear factor erythroid 2 (NF-E2) (G), early growth response protein 1 (EGR1) (H), and cellular oncogene fos (FOS) (I) (n = 3 per group). (J) Representative immunofluorescence images of toll-like receptor 2 (TLR2) fluorescently triple-stained with p-STING and p-NF-?B, respectively, on bone marrow (BM) cells after 12 days of treatment with oxymatrine in a mouse model of radiation thrombocytopenia. Blue for the 4?,6-diamidino-2-phenylindole (DAPI), purple for the TLR2, red for the p-STING, and green for the p-NF-?B. (K, L) Representative images of immunofluorescence of the nuclear translocation of NF-E2 (K) and RUNX1 (L) in Meg-01 cells after the treatment of oxymatrine for five days. Excitation wavelengths: 470 nm for NF-E2 and RUNX1 and 405 nm for DAPI. Data represent the mean ± standard deviation (SD) of three independent experiments. ?P < 0.05, ??P < 0.01, and ???P < 0.001 vs. the control group. EGFR: epidermal growth factor receptor; IL-17: interleukin 17; VEGF: vascular endothelial growth factor; TNF: tumor necrosis factor; JAK-STAT: janus kinase-signal transducer and activator of transcription; PI3K-Akt: phospholnositide-3 kinase-threonine kinase B; MAPK: mitogen-activated protein kinase; FoxO: forkhead box protein O; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. |