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. 2012 Dec 7;7(12):e50442. doi: 10.1371/journal.pone.0050442

Figure 5. HCN1 and HCN2 dependence on NT3.

Figure 5

A to D: DRG neurons cultured for 2 days in absence (A and B) or presence (C and D) of NT3 (40 ng/ml). HCN1 (A and C) and HCN2 (B and D) immunofluorescence examples are shown. Immunostaining for both was much reduced in the absence of NT3. Arrows in A and B show neurons visible under interference contrast; many were so weakly stained they were not/weakly visible under fluorescence. Note that in B, the left hand image includes smaller neurons; the right image includes a medium-sized and a large neuron. In C and D (with NT3), staining was stronger at the cell perimeter of medium to large neurons for both HCN1 and HCN2. Some smaller neurons showed elevated HCN2 staining (D). E and F: Quantitation of the effect of NT3 (40 ng/ml) on HCN1 and HCN2 edge staining after 2 days in culture. Medians for HCN1 and HCN2-edge immunostaining were both significantly greater (Mann Whitney test) for medium and large neurons in the presence of NT3. For HCN1 only, there was also a significant elevation in edge staining in small neurons with NT3. * P<0.05; **P<0.01, ***P<0.001, ****P<0.0001.