Journal: Frontiers in Pharmacology

Article Title: Regulating mitochondrial oxidative phosphorylation and MAPK signaling: wedelolactone as a novel therapeutic for radiation-induced thrombocytopenia

doi: 10.3389/fphar.2025.1508215

Figure Lengend Snippet: MAPK signaling pathway is necessary for WED-induced megakaryocyte maturation. (A-F) Western blot analysis showing the expression levels of RAS, p-MEK, p-ERK, FOS, NF-E2, and GATA1 in cells exposed to WED (2.5, 5, and 10 μM). *p < 0.05, **p < 0.01, ***p < 0.001 vs. the control group. (G) Flow cytometry analysis, with histograms displaying the proportion of CD41 + CD61 + cells. Data are displayed as mean ± SD (n = 3). ***p < 0.001 vs. the WED group.

Article Snippet: Add primary antibody and incubate overnight, then wash three times with PBST (PBS with 2% Tween-20), the secondary antibody was added and incubated at 37°C for 1 h. Antibodies against RAS (Abmart, T56672S), p-MEK1/2 (CST, 9154S), MEK1/2 (CST, 8727 S), p-ERK1/2 (Abmart, TA1015S), ERK1/2 (Abmart, TA0155), p-AMPK (Abmart, T55608F), AMPK (Abmart, T55326F), NF-E2 (Proteintech, United States, 11089-1-AP), GATA1 (Proteintech, United States, 10917-2-AP), FOS (Proteintech, United States, 66590-1-Ig), GAPDH (Proteintech, United States, 10494-1-AP).

Techniques: Western Blot, Expressing, Control, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Regulating mitochondrial oxidative phosphorylation and MAPK signaling: wedelolactone as a novel therapeutic for radiation-induced thrombocytopenia

doi: 10.3389/fphar.2025.1508215

Figure Lengend Snippet: Schematic model showing the role of WED in regulating megakaryocyte differentiation and platelet generation. WED promotes megakaryocyte differentiation and platelet generation by activating the MAPK signaling pathway (RAS/MEK/ERK1/2) and modulating oxidative phosphorylation via the AMPK signaling pathway. Activation of MAPK signaling results in the upregulation of key hematopoietic transcription factors, including FOS, NF-E2, and GATA1, which are essential for megakaryocyte differentiation and maturation. This process ultimately leads to the restoration of platelet counts, as demonstrated in the RIT mice model. MEP: Megakaryocyte-erythroid progenitor.

Article Snippet: Add primary antibody and incubate overnight, then wash three times with PBST (PBS with 2% Tween-20), the secondary antibody was added and incubated at 37°C for 1 h. Antibodies against RAS (Abmart, T56672S), p-MEK1/2 (CST, 9154S), MEK1/2 (CST, 8727 S), p-ERK1/2 (Abmart, TA1015S), ERK1/2 (Abmart, TA0155), p-AMPK (Abmart, T55608F), AMPK (Abmart, T55326F), NF-E2 (Proteintech, United States, 11089-1-AP), GATA1 (Proteintech, United States, 10917-2-AP), FOS (Proteintech, United States, 66590-1-Ig), GAPDH (Proteintech, United States, 10494-1-AP).

Techniques: Phospho-proteomics, Activation Assay