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. 2023 Apr 16;83(12):1953–1967. doi: 10.1158/0008-5472.CAN-22-3848

Figure 1.

Figure 1. Development of mutant Keap1 and Nfe2l2 alleles found in human lung cancer. A, The conditionally active (CA)-Keap1R554Q allele was generated by inserting a loxP-flanked, WT Keap1 cDNA containing exons 3–5 into intron 2 and introducing the R554Q mutation into endogenous exon 4 of the Keap1 gene. Before intranasal installation of adenoviral-Cre recombinase Keap1 is WT. Once the floxed cargo is excised, mutant Keap1R554Q is expressed. PA, poly A signal. B, The Lox-STOP-Lox (LSL)-Nfe2l2D29H allele was created by inserting a STOP cassette flanked by loxP sites into intron 1 and introducing the D29H mutation in endogenous exon 2 of the Nfe2l2 gene. Following Cre-mediated excision of the STOP cassette, mutant Nrf2D29H is expressed. C, Volcano plot of RNA-seq data from MEFs expressing Keap1R554Q/R554Q compared with Keap1+/+. N = 3, representative of two individual MEF lines. D, Volcano plot of RNA-seq data from MEFs expressing Nrf2D29H/D29H compared with Nrf2LSL/LSL, which lack Nrf2 expression. N = 3, representative of two individual MEF lines.

Development of mutant Keap1 and Nfe2l2 alleles found in human lung cancer. A, The conditionally active (CA)-Keap1R554Q allele was generated by inserting a loxP-flanked, WT Keap1 cDNA containing exons 3–5 into intron 2 and introducing the R554Q mutation into endogenous exon 4 of the Keap1 gene. Before intranasal installation of adenoviral-Cre recombinase Keap1 is WT. Once the floxed cargo is excised, mutant Keap1R554Q is expressed. PA, poly A signal. B, The Lox-STOP-Lox (LSL)-Nfe2l2D29H allele was created by inserting a STOP cassette flanked by loxP sites into intron 1 and introducing the D29H mutation in endogenous exon 2 of the Nfe2l2 gene. Following Cre-mediated excision of the STOP cassette, mutant Nrf2D29H is expressed. C, Volcano plot of RNA-seq data from MEFs expressing Keap1R554Q/R554Q compared with Keap1+/+. N = 3, representative of two individual MEF lines. D, Volcano plot of RNA-seq data from MEFs expressing Nrf2D29H/D29H compared with Nrf2LSL/LSL, which lack Nrf2 expression. N = 3, representative of two individual MEF lines.