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Fig. 2 | Cell & Bioscience

Fig. 2

From: Krüppel-like factor 7 deficiency causes autistic-like behavior in mice via regulating Clock gene

Fig. 2

Klf7 binds to Clock gene and forms a feedback loop with Clock gene. A Binding peaks for circadian rhythm genes, which were identified in two biological replicates by Chip-seq. B Luciferase reporter assay of Clock promoter in the presence of klf7. Klf7 positively regulates Clock gene transcription in a concentration dependent manner (n = 4 independent experiments). C Transcriptional analysis of Clock truncated promoter quantified by luciferase reporter assay. Specific 300 bp sequence of Clock promoter can be positively targeted by klf7 (n = 4 independent experiments). D PCR primers that can amplify the specific 300 bp sequence of Clock gene promoter in IP group (n = 4 independent experiments). E Clock gene inhibits klf7 expression in turn (n = 4). F Clock mRNA levels measured by qPCR in NC group, shklf7 group, NC and Clock group, shklf7 and Clock group (n = 4). G klf7 mRNA levels measured by qPCR in NC group, shklf7 group, NC and Clock group, shklf7 and Clock group (n = 4). H Protein levels of klf7 measured by western blot. I mRNA levels of Per2, Per3 and Rorb measured by qPCR in NC group, shklf7 group, NC and Clock group, shklf7 and Clock group (n = 4). The data are presented as the mean ± SEM. Statistical analysis (P*** < 0.001 and P**** < 0.0001) among empty vector group, klf7 (500 ng) group and klf7 (1000 ng) group in (B) were performed by one-way ANOVA test. Statistical analysis (P**** < 0.0001) among empty vector group, 280 bp group and 300 bp group in (C) were performed by one-way ANOVA test. Statistical analysis (P*** < 0.001, P**** < 0.0001 and ns: no significance) among NC group, 250 ng/μL group, 1000 ng/μL group, 2000 ng/μL group, and 4000 ng/μL group in (E) were performed by one-way ANOVA test. Statistical analysis (P* < 0.05, P** < 0.01, P*** < 0.001 and ns: no significance) among NC group, shklf7 group, NC + clock group and shklf7 + clock group in (F, G) and (I) were performed by one-way ANOVA test

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