Fig. 1

Species comparison of NOS2 mRNA, iNOS protein, and nitric oxide production. Microglia were unstimulated (CTL) or stimulated with IFN-γ plus TNF-α (I + T), IL-4, or IL-10 for 24 h. a NOS2 mRNA expression (mRNA counts/200 ng total RNA) was determined by NanoString. mRNA counts for each gene were normalized to two housekeeping genes (described in Methods) and are shown as mean ± SEM (n = 4–6 individual cultures), plotted on the same Y-axis scale. b Two representative Western blots of iNOS protein, with both species on the same gel. For each example, the full membrane was stained with Coomassie blue (lower panel), which was used to normalize iNOS protein levels. c Summary of fold changes in iNOS protein (mean ± SEM; n = 4–6 individual cultures for mouse and 22 for rat). For each Western blot, each iNOS band was normalized to total protein in that lane and then, iNOS levels for each treatment were normalized to unstimulated (control) microglia. d Nitric oxide production was monitored using the Griess assay (mean ± SEM; n = 6–11 individual cultures). Significant differences from unstimulated (control) cells are indicated: ***p < 0.001; ****p < 0.0001