Fig. 4

The majority of HCN4-IR cells do not express VGAT. a Schematic diagram of the recombinant allele in the VGAT-Venus BAC transgenic mouse. b Colocalization of HCN4-immunoreactivity and VGAT-Venus. b-1 Immunoreactivity for HCN4 (red) and NeuN (cyan). b-2 VGAT-Venus (green) and NeuN (cyan). b-3 Overlay of HCN4-immunoreactivity (red) and VGAT-Venus (green). b-4 PKCγ (gray). HCN4-IR cells indicated by arrowheads do not colocalize with Venus. c Colocalization of HCN4 (red), VGAT (green), and PV (cyan). c-1 Immunoreactivity for HCN4 (red). c-2 VGAT-Venus (green). c-3 Immunoreactivity for PV (cyan). c-4 Overlay of HCN4-immunoreactivity (red), VGAT-Venus (green), and PV-immunoreactivity (cyan). Arrowheads indicate HCN4- and PV-IR cells. Note that these cells do not colocalize with VGAT-Venus. It should be also noted that the intensity of PV-immunoreactivity in PV-IR and Venus-positive cells are higher than that in HCN4- and PV-IR cells. b, c Scale bars, 40 μm. The image was obtained in the same preparation shown in Fig. 3a, b. d Proportions of NeuN-IR cells that coexpress VGAT-Venus in each lamina. e Proportions of NeuN-IR cells that coexpress HCN4-immunoreactivity in each lamina. f Proportions of HCN4-IR cells that coexpress Venus in each lamina. g Venn diagrams showing the relationships among populations of HCN4-IR cells (red circle), VGAT-Venus-expressing cells (green circle), and NeuN-IR cells (gray rectangle) in each lamina. The number shown in each category indicates the average number of cells identified in an 8-μm thick slice