Figure 1.
Effect of autophagy deficiency on long-term hematopoiesis. (A-B) After the administration of poly:IC, the number of total BM cell (A) and frequency of LSK (Lineage−, Sca1+, c-Kit+) cells (B) were analyzed. Data are presented as means ± standard deviation (SD; n = 3 in each condition, 2 independent experiments; *P < .05 by Student t test). (C) Frequency of myeloid cells (CD11b+ and Gr1+) were examined in PBMCs derived from Atg7fl/fl (fl = flox) or Atg7fl/fl; Mx1-Cre mice. Histograms represent the fluorescent intensity of CD11b and Gr1 antibody, and the graph shows frequency of CD11b+ and Gr1+ cells. Data are presented as means ± SD (n = 3 in each condition, 2 independent experiments; *P < .05 by Student t test). (D) Photograph of spleen and weigh of spleen were measured in Atg7fl/fl or Atg7fl/fl; Mx1-Cre mice. Graph shows weight of spleen. Data are presented as means ± SD (n = 3 in each condition, 2 independent experiments; *P < .05 by Student t test). (E-F) ΔΨm (E) and mitochondrial superoxide level (F) were examined in HSCs derived from Atg7fl/fl or Atg7fl/fl; Mx1-Cre HSCs. Histograms represent the fluorescent intensity of indicated dyes, and graphs show relative values against Atg7fl/fl-derived HSCs. Data are presented as means ± SD (n = 3 in each condition, 2 independent experiments; *P < .05 by Student t test). (G-I) After the administration of poly:IC, Atg7fl/fl; Mx1-Cre HSCs (Ly5.2) were transplanted into lethally irradiated recipient mice (Ly5.1) along with wild-type whole BM cells (Ly5.1) (G). (H) After 4, 12, and 20 weeks from primary transplantation, peripheral blood (PB) of each recipient was analyzed. (I) Moreover, PB was analyzed at 20 weeks after chimeric whole BM cells derived from recipient mice were serially transplanted. Graphs show the chimerism of PB in each recipient. Data are presented as means ± SD (n = 9, 2 independent experiments; *P < .05 by Student t test). Black and white bars represent Atg7fl/fl (used as a control) and Atg7fl/fl; Mx1-Cre-derived cells, respectively.