Fig. 1: Highly efficient KO generation in primary human resting CD4+ T cells.
a, Schematic overview of the pipeline to establish polyclonal KOs in human resting CD4+ T cells. b, Viability of CD46 KO or untreated WT resting CD4+ T cells kept in culture for up to 6 weeks. Cell viability was assessed at the indicated time points by flow cytometry. Mean ± s.e.m. are shown (n = 3). c, T-cell proliferation assay. CFSE-labeled CD46 KO and WT CD4+ T cells analyzed by flow cytometry 1 week after nucleofection. Anti-CD3/CD28 antibody-activated CD4+ T cells (activated) served as positive control (Ctrl). One representative experiment is shown (n = 3). d, WT and CD46 KO resting CD4+ T cells were analyzed by flow cytometry for expression of T-cell activation markers CD25 and CD69. Mean ± s.e.m. are shown (n = 3). e, Resting CD4+ T cells (WT or CD46 KO) were analyzed for cell surface expression of CD46 2 weeks after nucleofection. Shown are dot plots of flow cytometric analyses for cells from three individual donors and WT cells from one of these donors. Fig. 1a created with BioRender.com.
