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. 2023 May 3;38(13):959–973. doi: 10.1089/ars.2022.0058

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FIG. 3. — Analysis of control versus Keap1-KO CD4+ T cells. (A) Sustained increase of mRNA levels of Nrf2 target genes Nqo1, Hmox1, Gclc, and Gclm in Keap1-KO CD4+ T cells compared with control cells 96 h after electroporation. (B) Immunoblotting of control and Keap1-edited CD4+ T cells 96 h after electroporation and before adoptive transfer for KEAP1 with 73% knockdown and NQO1 with 50% increase of expression. (C) Comparable FoxP3 and activation marker expression and (D) comparable expression of TNFα, IL17, and IL10 72 h after electroporation. (E) Panel to test for T cell exhaustion markers showed no changes of inhibitory receptor expression 96 h after electroporation; n = 3 in each group in (A), (C), (D), and (E); n = 2 in each group in (B). Statistical analyses were conducted using paired, two-tailed T-test. **p < 0.001. Gclc, glutamate-cysteine ligase catalytic subunit; Gclm, glutamate-cysteine ligase modifier subunit; Hmox1, heme oxygenase 1; Nqo1, NAD(P)H quinone dehydrogenase 1.

FIG. 3. — Analysis of control versus Keap1-KO CD4+ T cells. (A) Sustained increase of mRNA levels of Nrf2 target genes Nqo1, Hmox1, Gclc, and Gclm in Keap1-KO CD4+ T cells compared with control cells 96 h after electroporation. (B) Immunoblotting of control and Keap1-edited CD4+ T cells 96 h after electroporation and before adoptive transfer for KEAP1 with 73% knockdown and NQO1 with 50% increase of expression. (C) Comparable FoxP3 and activation marker expression and (D) comparable expression of TNFα, IL17, and IL10 72 h after electroporation. (E) Panel to test for T cell exhaustion markers showed no changes of inhibitory receptor expression 96 h after electroporation; n = 3 in each group in (A), (C), (D), and (E); n = 2 in each group in (B). Statistical analyses were conducted using paired, two-tailed T-test. **p < 0.001. Gclc, glutamate-cysteine ligase catalytic subunit; Gclm, glutamate-cysteine ligase modifier subunit; Hmox1, heme oxygenase 1; Nqo1, NAD(P)H quinone dehydrogenase 1.