Fig. 1
MK C3G promotes hematopoietic recovery following 5-FU treatment by enhancing megakaryocyte (MK) maturation (A) Scheme of 5-FU-induced myeloablation. Blood drops indicate the days of blood extraction. Femurs indicate the days of femur and BM harvest. Platelet count in (B) tgC3GPf4 and (C) C3GPf4-KO mice and their controls. Evaluation of (D, G) WBC, (E, H) RBC, and (F, I) hemoglobin in (D–F) tgC3GPf4 and (G–I) C3GPf4-KO and their controls. Data (mean ± SEM) correspond to two independent experiments. wtC3GPf4, n = 4; tgC3GPf4, n = 5; C3GPf4-wt, n = 5; C3GPf4-KO, n = 5. (J, L) Graphics showing the mean ± SEM of MK count in the first 5 mm of femoral BM from the growth plate, at the indicated days after 5-FU, in (J) wtC3GPf4 and tgC3GPf4 and in (L) C3GPf4-wt and C3GPf4-KO. (K, M) Representative photomicrographs of BM sections stained with H&E. Arrowheads indicate MKs. The insets show a 2.5X magnification of the regions highlighted in the images. Scale bar: 0.25 mm. (N, O) Normalized counts (mean ± SEM) of mature MKs (CD42+CD61+ cells) by flow cytometry in BM extracts from (N) tgC3GPf4, and (O) C3GPf4-KO mice and their controls at the indicated days after 5-FU. (P, Q) Violin plots show the median size of MKs in the inner part of the diaphysis. (R, S) MK (mean ± SEM) quantification along (R) wtC3GPf4 and tgC3GPf4 femora 7 days after 5-FU injection, and (S) C3GPf4-wt and C3GPf4-KO femora 10 days after 5-FU injection. Two to three independent experiments were performed per day of analysis. wtC3GPf4, n = 4–6; tgC3GPf4, n = 4–8; C3GPf4-wt, n = 4–6; C3GPf4-KO, n = 4–6. p-values in (B–I, J, L, R, S) were calculated using two-way ANOVA with uncorrected Fisher’s LSD post-hoc test. Unpaired t-test was used in (N–Q). *p ≤ 0.05. **p ≤ 0.01. ***p ≤ 0.001