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. 2022 Sep 7;23(18):10342. doi: 10.3390/ijms231810342

Figure 5.

Figure 5

Deficiency of Keap1 and its isoforms α and β lead to gene dysregulation of cell behaviour, cycle, and apoptosis. (A) Real-time qPCR was used to quantify the mRNA expression levels of Keap1, CTNNA1, CTNNB1, SNAI1, SNAI2, CDH1, CDH2, FN1, MMP17, MMP9, and Vimentin in Keap1+/+, Keap1−/−, and Keap1β(Keap1Δ1–31) cell lines. (B) Western blotting was employed to determine the protein abundances of Keap1, CTNNA1, CTNNB1, SNAI1, SNAI2, CDH1, CDH2, FN1, MMP17, MMP9, and Vimentin in Keap1+/+, Keap1−/−, and Keap1β(Keap1Δ1–31) cell lines. (C) The mRNA expression levels of Keap1, CTNNA1, CTNNB1, SNAI1, SNAI2, CDH1, CDH2, FN1, MMP17, MMP9, and Vimentin in Keap1−/−, Keap1-Restored and Keap1α-Restored cell lines were determined by real-time qPCR. (D) The protein abundances of Keap1, CTNNA1, CTNNB1, SNAI1, SNAI2, CDH1, CDH2, FN1, MMP17, MMP9, and Vimentin in Keap1−/−, Keap1-Restored, and Keap1α-Restored cell lines were visualized by Western blotting with indicated antibodies. (E) The mRNA expression levels of genes controlling cell cycle, i.e., P53, P21, CDK2, CDK6, and CyclinD1 in Keap1+/+, Keap1−/−, and Keap1β(Keap1Δ1–31) cell lines were quantified by real-time qPCR. (F) The mRNA levels of genes controlling cell apoptosis, i.e., CASP3, CASP4, CASP6, BCL2, and BCL2L1 in Keap1+/+, Keap1−/−, and Keap1β(Keap1Δ1–31) cell lines were evaluated by real-time qPCR. (G) Real-time qPCR was also subjected to determining the mRNA expression levels of P53, P21, CDK2, CDK6, and CyclinD1 in Keap1−/−, Keap1-Restored, and Keap1α-Restored cell lines. (H) The mRNA expressions of CASP3, CASP4, CASP6, BCL2, and BCL2L1 in Keap1−/−, Keap1-Restored, and Keap1α-Restored cell lines were also measured. Statistically significant increases ($) or decreases (*) were indicated by p ≤ 0.01 (n ≥ 3, or =3 × 3).