Figure 4. Miz1 is a transcriptional activator of genes involved in vesicular transport.

(a) Miz1 activates the Rorc and Ambra1 promoters and the POZ domain is required for activation. HEK293 cells were transiently transfected with plasmids containing promoter sequences of Ambra1 or Rorc in front of luciferase and with expression vectors encoding Miz1 or Miz^ΔPOZ. Empty vectors were used as control. Values were normalized with CMV-βGal. Error bars are standard deviation (SD) of biological triplicates.

(b) Direct target genes of Miz1 are downregulated in cerebella of Miz1^ΔPOZNes mice. Microarray analysis was performed from cerebella of mice at the indicated ages. The panels show box plots of relative gene expression (plotted as log₂ fold change) of the 140 Miz1 direct target genes comparing control and Miz1^ΔPOZNes mice. The p-value was calculated using a Mann–Whitney–Wilcoxon-test. Microarrays were performed as technical duplicates from RNA pooled from two animals for each genotype.

(c) Gene set enrichment analysis ¹⁸ documenting significant downregulation of Miz1 target genes in cerebella of Miz1^ΔPOZNes mice. NES: normalized enrichment score (NES). Target genes of Miz1 were the only significant gene set in this analysis.

(d) qRT-PCR documenting expression of selected Miz1 target genes in cerebella of control and of Miz1^ΔPOZNes mice at the indicated ages. Data represent the mean of two independent biological experiments each with three technical replicates. Error bars are SEM.