Extended Data Fig. 10: Staining of blood platelets with Thiazole Orange after Cyclophosphamide treatment, and fate map** of Flt3Cre^tg/+ R26^Tom/+ mice after 5FU and antibody-induced platelet depletion.

a, Representative flow cytometry profiles (% of parent gates) of Tomato labelling in Thiazole Orange positive (TO⁺) platelets of a Flt3Cre^tg/+ R26^Tom/+ mouse before CP (upper panels), and on day 4 (middle panels) and 45 (lower panels) after CP. b, Representative flow cytometry profile (% of parent gates) of TO labelling in Tom⁺ and Tom⁻ platelets of a Flt3Cre^tg/+ R26^Tom/+ mouse before and after CP. c, Ratio (mean ± s.e.m.) between TO⁺ Tom⁻ and TO⁺ Tom⁺ platelets in Flt3Cre^tg/+ R26^Tom/+ mice before and after CP (day 0). Analysis at baseline (day -3, n = 16), and on day 4 (n = 10), 7 (n = 10), 18 (n = 10), and 45 (n = 7). Compared to baseline, ****P = 3.77 × 10⁻⁷ for d4 and ****P = 3.90 × 10⁻⁵ for d7. Two-tailed Mann-Whitney test. d, Blood cell counts, relative to baseline (d-2), of 8–10 week old Flt3Cre^tg/+ R26^{Tom /+} mice before and after 5-Fluorouracil (5FU, n = 3) treatment. Lines connect the means for each time point. e, Flt3Cre-tdTomato labelling of BM LIN⁻FLT3⁺ progenitor cells in Flt3Cre^tg/+ R26^{Tom /+} mice, untreated (n = 4) and 5–10 days after 5FU treatment (n = 2). Dots represent individual mice and lines represent means. f, Blood cell counts, relative to baseline of 12–13 week old Flt3Cre^tg/+ R26^Tom/+ mice (n = 3), each analyzed before treatment (baseline; d-10 or -3) and on day 3, 7 and 11 after treatment with anti-CD42b antibody (αCD42b). Lines connect the means for each time point. g, BM cellularity (left) and BM LK CD150⁺CD41⁺ MkP (right) cell number recovered after crushing both femurs, tibiaes, and pelvic bones of 12–13 week old untreated (n = 5) and αCD42b-treated (n = 4) mice. Dots represent individual mice and lines represent mean ± s.e.m. BM cellularity was not significantly different, whereas BM MkP cell number was significantly increased after αCD42b treatment (**P = 1.59 × 10⁻²); two-tailed Mann-Whitney test.

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