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. Author manuscript; available in PMC: 2015 Oct 2.
Published in final edited form as: Nature. 2015 Jan 19;520(7545):94–98. doi: 10.1038/nature14051

Fig. ED4. Quantitation of MC4R and Kir7.1 RNA in PVN cells.

Fig. ED4

Single molecule RNA detection in sections was quantitated by counting fluorescent dots associated with individual cells (Fig. ED3). Background threshold was determined from the number of dots per cell in sections resulting from hybridization using a negative bacterial DNA control, or from hybridization of the MC4R probe to sections from the MC4R knockout mouse (a, columns 1 and 2). Threshold-subtracted dot numbers were then used to determine the % of PVN cells expressing MC4R or Kir7.1, and the % of MC4R cells expressing Kir7.1; cells were considered positive if the number of dots exceeded the mean of the negative controls by 3x std (b).