Figure 8. Individual γ-Pcdh isoforms can differentially regulate Wnt signalling in the cerebral cortex in vivo.

(a) Schematic of transgenic mouse alleles utilized. The Wnt reporter allele (top⁶⁸) has 6 TCF/LEF Wnt-responsive binding sites and an hsp68 minimal promoter driving histone 2B-GFP. The A1-mCherry and C3-mCherry transgenes²⁹,³² are inserted at the ubiquitous Rosa locus. The transgenes are driven by Rosa and CAG elements (RC) only after a floxed stop cassette is excised by Cre. Yellow triangles, loxP sites. (b,c) Cortical lysates from animals overexpressing γ-Pcdh-A1 or –C3 were used in western blots and probed for GFP to visualize changes in H2B-GFP level, reflecting Wnt signalling activity. Representative Western blot shows an increase in H2B-GFP band intensity in A1-overexpressing animals, while C3-overexpressing animals exhibit a decrease in H2B-GFP band intensity. Blots were probed with anti-mCherry antibody to verify expression of exogenous A1 and C3 isoforms, while β-tubulin is used as a loading control. (d) Band intensities of H2B-GFP were normalized to those of β-tubulin in the same lane. Means ± SEM of ≥5 animals were graphed and analysed using one-way ANOVA with Bonferroni post hoc test (to correct for multiple comparisons). *p < 0.05, **p < 0.01. MW, molecular weight; kDa, kilodaltons.