Fig 1: NRF2-dependent regulation of genes in autophagy-deficient ameloblasts. (A) Quantitative reverse transcription polymerase chain reaction (RT-PCR) analyses for the indicated genes in mHat9d cells treated with a control (blue bars) and Nrf2 overexpression (red bars) vector. ***P < 0.001. n = 6 per group. (B) Immunoblotting for the indicated molecules in wild-type (WT), Atg7 knockout (KO), and Atg3 KO mHAT9d cells. (C) Transmission electron microscopy in WT, Atg7 KO, and Atg3 KO mHAT9d cells. Yellow arrows indicate the autophagosomes. Red arrow indicates vesicle-like abnormal structures. Scale bars: 2 μm. (D) Quantitative RT-PCR analyses for the indicated genes in WT and Atg7 KO mHAT9d cells (upper panel) and WT and Atg3 KO mHAT9d cells (lower panel) with and without Nrf2 knockdown (KD). **P < 0.005. ***P < 0.001. n = 6 per group.
Fig 2: Identification of genes with compromised expression in the teeth of autophagy-deficient mice. (A) Schematic diagram of binding sites (BSs) for NRF2 in the promoter region (10 kb upstream from transcription start site) of each gene related to amelogenesis imperfecta. The conserved NRF2 BSs among 8 species were selected for experimental validation. (B) Quantitative reverse transcription polymerase chain reaction analyses for the indicated genes of upper incisors from wild-type (blue bars) and Atg7 cKO (green bars) mice. ***P < 0.001. n = 6 per group. (C) Chromatin immunoprecipitation analyses for each BS in Dlx3, Klk4, Nectin1, Bcl11c, Pax9, and Ltbp3. ***P < 0.001; ns, not significant. n = 6 per group.
Supplier Page from Sino Biological, Inc. for Mouse Nrf2/NFE2L2 Gene ORF cDNA clone in cloning vector