Fig. 2
Laminar distribution of HCN4-IR cells. a Immunohistochemistry of the SDH in an HCN4+/tTA_TRE mouse. Left panel, HCN4 immunoreactivity (red). Center panel, PKCγ immunoreactivity (green). Right panel, high-magnification image expanding the white box of the center panel. Overlay of HCN4 (red), PKCγ (green), and NeuN (cyan) IR signals. HCN4-IR cells are indicated by arrowheads. Dashed lines indicate the borders of the PKCγ plexus (upper line, border between lamina IIid and lamina IIiv; lower line, border between lamina IIiv and lamina III). b Immunohistochemistry of the SDH in an HCN4Luc/tTA_TRE DOX(+) mouse. Left panel, no HCN4-IR cells can be identified. Center panel, the pattern of PKCγ-immunoreactivity is essentially the same as in a. Right panel, colors and lines correspond to those in a. Note that there are no HCN4-IR cells. a, b Scale bars, 200 μm (left and center panels), 40 μm (right panel). c Percentages of HCN4-IR cells per NeuN-IR cells in each lamina. d Proportion of HCN4-IR cells per each lamina I–III. The ratio was calculated as the number of HCN4-IR cells in a layer divided by the total number of HCN4-IR cells in laminae I–III