Fig. 2: Loss of TNFR2 expression does not affect MDSC accumulation in male mice.

Livers were resected at the indicated time intervals postintrasplenic/portal injection of 5 × 10⁵ GFP-tagged MC-38 cells (green) (a, c) or 5 × 10⁴ MC-38 cells (b), 14 μm frozen sections prepared and immunostained with the indicated antibodies followed by Alexa Fluor 568 (red) for Gr-1, and Alexa Fluor 647 (cyan) or Alexa Fluor 700 (white) for CD11b and Alexa Fluor 647 (cyan) for TNFR2. Shown in a (left) are representative images of 20 fields acquired for each experimental group (n = 3, day 9) and in a (right) the mean numbers (±s.e.m.) of CD11b⁺Gr-1⁺ cells enumerated in each group at the indicated time intervals following tumor injection. Shown in b are results of FC performed on immune cells isolated 9 days post tumor injection and immunostained with the indicated antibodies. They are expressed as means (±s.e.m.) of three separate analyses, each performed on livers obtained from six mice per group. Shown in c (left) are representative images of TNFR2⁺CD11b⁺Gr-1⁺ cells visualized in 30 fields in sections derived from male mice inoculated with MC-38 cells 9 days earlier and in c (right) the results expressed as % TNFR2⁺ cells (±s.e.m.) of total CD11b⁺Gr-1⁺ cells counted per field in sections derived at the indicated time intervals following tumor injection. NS, not significant as determined by the a two-tailed Student's t-test. Scale bars correspond to 100 µm on histology images.