Neuronal COUP-TFII Regulation Figure 2. COUP-TFII mRNA levels in the hypothalamus of eGHI clamped mice. (A) Blood glucose and plasma insulin (B-C) RT-qPCR analysis of mRNA levels in the hypothalamus of eGHI clamped mice. (B) Left panel: POMC mRNA levels; middle panel: AgRP mRNA levels; right panel: NPY mRNA levels (C) COUP- TFII mRNA levels. Saline control mice (%), eGHI (&) Data are expressed as mean 6 S.E.M. (n = 6 mice/group). **P#0.01 when compared to saline control mice. doi:10.1371/journal.pone.0013464.g002 staining for both proteins. A large number of neurons expressing the SF1 protein were observed in the VMH (figure 5A, red fluorescence). Interestingly, not all VMH cells were expressing COUP-TFII (figure 5B, green fluorescence). By overlapping the two fluorescences, we observed that all the COUP-TFII positive neurons were also expressing the SF1 protein (overlapping green and red fluorescence) (figure 5E, yellow fluorescence). However, some SF1 cells were not expressing COUP-TFII (white arrows), suggesting that we had at least two distinct cellular subpopulations of SF1 neurons within the VMH, the COUP-TFII positive and the PLoS ONE | www.plosone.org 4 October 2010 | Volume 5 | Issue 10 | e13464