Figure 2.

Basal levels of autophagy are necessary for B-cell survival in the periphery. Spleen cells from wild-type C57BL/6 mice (B6), Atg5^f/+ CD21cre (littermates CD21 cre) or Atg5^f/+ Mb1cre (littermates Mb1 cre), Atg5^f/− CD21 cre (CD21 cre) or Atg5^f/− CD21 cre (Mb1 cre) were stained with anti-B220 and anti-CD19 Abs. (a and b) Representative dot plots obtained after flow-cytometry analysis are depicted in (a) showing the percentages of B220⁺CD19⁺ B cells. (b) Individual values for each mouse tested of absolute numbers of B and T cells, in the spleen obtained from B6 mice n=16, littermate CD21 cre mice n=10, littermate Mb1 cre mice n=9, CD21 cre mice n=15, and Mb1 cre mice n=9. Means and standard deviations (S.D.) are indicated. (c–e) Expression was assessed for the spleen of the same animals for the expression of (c) IgD, IgM and (d) CD21 and CD23. Representative dot plots after flow-cytometry analysis show the percentages among B220⁺ cells of mature B cells (IgM^lowIgD^hi), transitional 1 and 2 (T1 and T2, respectively, IgM^hiIgD^lo and IgM^hiIgD^hi), follicular (FO) B cells (CD21⁺CD23^hi), and marginal zone (MZ) B cells (CD21^hiCD23^lo). (e) Individual values for each mouse tested of absolute numbers different B-cell populations, in the spleen obtained from B6 mice n=16, littermate CD21 cre mice n=10, littermate Mb1 cre mice n=9, CD21 cre mice n=15, and Mb1 cre mice n=9. Means and S.D. are indicated. (f and g) Peritoneal lavage was also performed on some animals and the cells obtained were stained by anti-CD5 and anti-B220 Abs. Representative histogram obtained after analysis of cells from the peritoneal lavage by flow cytometry, with the different populations indicated: B-2 B cells (B220^hiCD5⁻), B-1a B cells (CD5⁺B220^lo), and B-1b B cells (CD5⁻B220^lo). (g) Individual values for each mouse tested of percentages of different B-cell populations, in spleens obtained from B6 mice n=17, littermate CD21 cre mice n=8, littermate Mb1 cre mice n=4, CD21 cre mice n=15, and Mb1 cre mice n=6. *P<0.05, **P<0.01, ***P<0.001 (Mann–Whitney U-test). Hi, high; lo, low