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Fig. 2 | Cell Communication and Signaling

Fig. 2

From: Targeting the orphan nuclear receptor NR2F6 in T cells primes tumors for immune checkpoint therapy

Fig. 2

CRISPR/Cas9 mediated gene knockout in primary mouse T cells. a Schematic overview of CRISPR/Cas9 mediated gene knockout in isolated CD4+ T cells from Cas9 transgenic mice. b 10 days post-treatment, flow cytometry assays were performed to measure the loss of CD44 or CD69 in CD4+ Cas9 transgenic T cells targeted with sgRNAs against CD44 or CD69. c Efficient gene deletion achieved in primary T cells cells following treatment with different sgRNAs. Knockout efficiencies were calculated based on surface marker expression in comparison to NTC treated cells (CD44 KO: d6 p = 0.0002, d10 p = 0.0009, d13 p = 0.0026, CD69 KO: d6 p = 0.0003, d10 p = 0.0009, d13 p = 0.0062). d FACS plots and e quantification of CD4+ T cells with NTC or Nr2f6 CRISPR/Cas9 mediated knockout on day 10, re-stimulated with PdBU/Ionomycin for 4 h showing enhanced IFNγ cytokine production with Nr2f6 loss compared to NTC control cells (p = 0.0429). NTC, non-targeting control, sgRNA, single guide RNA, Cas9 Tg, Cas9 transgenic. The above experiments are repeated at least two times with similar results. Error bars represent the mean ± SEM

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