Fig 2: H2S promotes megakaryocytes/platelets generation but does not improve survival in TPO-/- mice. a Without radiation exposure, NaHS treatment (20 µmol kg-1 day-1) for 14 days caused an increase in circulating platelet levels in both WT and TPO-/- mice. b NaHS treatment (100 µmol L-1) increased the number of megakaryocytes identified as CD61-positive cells using flow cytometry in cultured fetal liver cells isolated from TPO-/- mice. c, d NaHS treatment (100 µmol kg-1 day-1) increased circulating platelet levels on day 7 after radiation exposure in both WT and TPO-/- mice (c). However, the effects of NaHS treatment (100 µmol kg-1 day-1) in increasing circulating platelet levels were blunted in TPO-/-mice (d). e Morphological examination of the bone marrow showed that NaHS treatment (100 µmol kg-1 day-1) did not increase the number of megakaryocytes (black arrows) in the TPO-/- mice exposed to radiation. Scale bar = 250 µm. f Bleeding was not ameliorated with NaHS treatment (100 µmol kg-1 day-1) in the TPO-/- mice exposed to radiation at day 7. g Likewise, fecal occult blood was not ameliorated with NaHS treatment (100 µmol kg-1 day-1) in the TPO-/- mice exposed to radiation at day 7. h Survival was not improved with NaHS treatment (100 µmol kg-1 day-1) in TPO-/- mice exposed to radiation. **P < 0.01 versus vehicle-treated mice exposed to radiation, log-rank test. Data in the graphs are means ± SEM. P values less than 0.05 represent statistical significance

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: Upregulation of BGN in BCSCs derived from PyMT tumor cells. (A) Venn diagram depicting the upregulated genes in CD29hi CD61+ (P1) and ALDH+ (P2) BCSCs as well as overlapping genes in both BCSCs as compared to bulk cells (P0). (B) KEGG pathway analysis showed that ECM receptor interaction and focal adhesion pathways were commonly upregulated in both BCSCs. (C) Heat map data showing the top 40 candidates identified from RNA sequencing. qRT-PCR data showing validation of genes in BCSCs, CD29hi CD61+ (P1) and ALDH+ (P2) as compared to bulk cells, (D) expression of ECM genes, (E) BCSCs marker expression, (F) ECM genes upregulated in BCSCs. Immunoblots depicting expression of BGN in sorted BCSCs from MMTV-PyMT tumor cells (FF99), (G) ALDH+ and (H) CD29hi CD61+ compared to bulk cells. Data shown as mean ± SEM of three independent experiments. Statistical significance was determined using two-tailed t-test; * p = 0.05 as compared to bulk cells.

Fig 5: p18 IL-1a Drives Rapid Thrombopoiesis and Wound Healing, and Is Generated during Sepsis in Humans(A and B) Platelet count by flow cytometry in mice under basal conditions (A), or over time after anti-CD42b-mediated platelet depletion (B).(C) Flow cytometry for the number of CD41+ and CD61+ MKs in the bone marrow of control and IL-1a TM mice under basal conditions.(D) Cleaved IL-1a-specific ELISA reporting the level of cleaved IL-1a within the serum of control mice treated ±dabigatran (+Dabi) and IL-1a TM mice over time after anti-CD42b-mediated platelet depletion.(E–G) Representative images showing gross healing of 4 mm excisional skin wounds (E), quantitation of wound area (F) over time, and rate of closure (G) in mice as indicated.(H–K) Representative images and quantitation of Ly6G+ neutrophils (H and I) or Mac3+ macrophages (J and K) recruited to the granulation tissue underlying wounds at the times indicated.(L) ELISA data showing release of cleaved IL-1a or IL-1ß from wounded skin.(M) Sandwich ELISA data showing reactivity of the p18-specific ELISA to p17, p18, or p33 IL-1a.(N and O) p18-specific ELISA data reporting level of p18 in plasma from control individuals or patients with sepsis-associated ARDS (N). Red circles indicate +VE microbiology in bronchoalveolar lavage fluid. (O) Cleaved IL-1a-specific ELISA reporting the level of cleaved IL-1a within the serum of control and IL-1a TM mice during LPS-induced endotoxemia. Data represent mean ± SEM; n = =4 (A, B, and D), n = 3 (C), n = =5 (F and G), n = =10 wounds (I and K), n = 20 wounds (L), n = 2 (M); p = *=0.05, **=0.01, ***=0.001; NS = not significant. See also Figure S4 and Table S2, S3.