Fig. 1

Identification of Klf9-regulated genes in HT22 cells by RNA-sequencing. a Treatment of HT22 [TR/TO-Klf9] cells with doxycycline (dox; 1 μg/ml) for 8 h increased Klf9 mRNA ~10 fold compared to vehicle treated cells, but had no effect in parent HT22 cells. The baseline Klf9 mRNA level did not differ between parent and [TR/TO-Klf9] line. Means with the same letter are not significantly different (p < .05; ANOVA followed by Tukey’s post-hoc test). b Dox-induced expression of Klf9 reduces luciferase activity from the pGL4.23-3xBTE plasmid, but not from pGL4.23-empty. The asterisk indicates a statistically significant difference by Student’s two-sample t-test (p < .05). c Validation by RTqPCR of four genes found to be repressed by Klf9 by RNA-seq. Treatment with dox for 8 h reduced mRNA levels for the four genes in HT22 [TR/TO-Klf9] but not in parent HT22 cells. The asterisks indicate statistically significant differences from parent cells treated with vehicle or dox, and from HT22 [TR/TO-Klf9] cells treated with vehicle (p < .05; ANOVA followed by Tukey’s post-hoc test). d Time-course showing induction of Klf9 mRNA following treatment of HT22 [TR/TO-Klf9] cells with dox. e Validation of repression (9 genes) or induction (3 genes) of Klf9 target genes after treatment of HT22 [TR/TO-Klf9] cells with dox for different times. The asterisks indicate statistically significant differences from the zero time point (p < .05; ANOVA followed by Tukey’s post-hoc test)