Colocalization of AVP and MIF in the PVN. Top, Low-power representative fluorescence images of the PVN showing endogenous MIF (green) and AVP (red) immunoreactivities and their colocalization (orange) at 3 different levels (−1.60, −1.88, and −2.12 mm relative to the bregma). Bars=100 μm. Middle, Representative fluorescence micrographs are a higher-power view (from the inset shown in the top left) showing MIF (green) and AVP (red) immunofluorescence at PVN level −1.60 mm relative to bregma. Examples of AVP and MIF colocalization are indicated by white arrows. Bar=20 μm. Bottom left, Bar graph showing the number of immunoreactive AVP and MIF cells counted at each of the 3 PVN levels and also the number of overlapping AVP- and MIF-positive cells. Data are mean±SEM of numbers of AVP-, MIF-, or AVP/MIF-positive cells (n=3 rats). Bottom right, Bar graph showing the colocalization of immunoreactive AVP and MIF in various subregions of the PVN at each of the levels investigated. dp indicates dorsal parvocellular; mp, medial parvocellular; pm, posterior magnocellular; vp, ventrolateral parvocellular; lp, lateral parvocellular. Data are mean±SEM of numbers of AVP/MIF-positive cells (n=3 rats).