Figure 1.
PKC knock-out mouse generation and immunostaining. A, Schematic illustration of the generation of PKCβ−/− mice. PKCβ gene has five exons, three of which (3, 4, and 5) are shown. Targeted embryonic stem (ES) cells (Prkcbtm1a(EUCOMM)Wtsi line EPD0233_5_F09) were obtained from The International Mouse Phenotyping Consortium and were injected into C57BL/6J blastocysts to generate chimeras. Chimeric mice were then bred with C57BL/6J to generate PKCβ targeted germline mice (PKCβ+/loxP). PKCβ+/loxP mice were bred with CMV-Cre mice (The Jackson Laboratory, 006054) to delete exon 4, generating PKCβ−/+ mice, which were used to establish the PKCβ−/− mouse line. Mouse genotypes were determined by PCR. B, Antibody staining of PKCα and vGluT1 in P9 WT and PKCα−/− calyces (“overlay”). C, Antibody staining of PKCβ and vGluT1 in P9 WT and PKCβ−/− calyces (“overlay”). D, Left, PKCα immunostaining staining intensity (FPKCα; a.u., arbitrary units; mean + SEM) in P7–P10 WT (14 calyces, 4 mice) and PKCα−/− calyces (15 calyces, 4 mice). **p < 0.01 (t test). Right, PKCβ immunostaining staining intensity (FPKCβ) in P7–P10 WT (24 calyces, 3 mice) and PKCβ−/− calyces (16 calyces, 3 mice). **p < 0.01 (t test).