Figure 5.

Detection of TNFR2 expression in islet cells. (A) Flow cytometry staining with anti-TNFR2 antibody and isotype control in islets isolated from 5 week old mice and cultured for 4–5 days. Auto-fluorescence in the FL1 channel was used to gate on the β cell population, and dead cells were excluded using 7AAD. The gated cells were negative for the hematopoetic cell marker CD45. Values given are the average %TNFR2+ cells +/- SEM (n = 4 each strain), gated on FL1+ cells and corrected for background isotype control staining. Results representative of 3 independent experiments. (B) Immunohistochemical staining of paraffin sections from TNFR2KO (i, iv), 12–14 week old NOD (ii, v) and Idd9 congenic (iii, vi) pancreas stained with antibodies to TNFR2 (i-iii) or with the secondary antibody alone (iv-vi). Pancreatic sections from 3 different Idd9 congenic and NOD mice, and 1 B6.TNFR2KO mouse, were examined.