Journal: Heliyon

Article Title: CRISPR/Cas9 editing reveals IRF8 regulated gene signatures restraining plasmablast differentiation

doi: 10.1016/j.heliyon.2023.e17527

Figure Lengend Snippet: Nucleofection of primary B cells with Cas9 and sgRNA targeting Ptprc . (A) Schematic of the Ptprc gene showing the locations of exons and sgRNAs. (B) Schematic of experimental design depicting the steps and time frames of primary B cell purification, pre-stimulation, nucleofection with Cas9/sgRNA, and analysis. (C) Histogram showing the efficiency of primary B cell nucleofection measured by GFP fluorescence. The Ptprc deletion efficiency as measured by B220 (D) and CD45 (CD45.2) (E) surface staining in cells nucleofected with each of the five sgRNAs depicted in A.

Article Snippet: For nucleofection experiments, splenic CD43 − B cells were isolated using the B-cell Isolation Kit (Miltenyi; 130-090-862).

Techniques: Purification, Fluorescence, Staining

Journal: Heliyon

Article Title: CRISPR/Cas9 editing reveals IRF8 regulated gene signatures restraining plasmablast differentiation

doi: 10.1016/j.heliyon.2023.e17527

Figure Lengend Snippet: Assessment of sgRNA targeting Irf8 in primary B cells. (A) Schematic of the Irf8 gene showing the locations of exons and sgRNAs. (B) Histogram showing the efficiency of primary B cell nucleofection measured by GFP fluorescence. (C) The sgRNA editing efficiency as measured by IRF8 intracellular staining in cells nucleofected with each of the five sgRNAs depicted in A.

Article Snippet: For nucleofection experiments, splenic CD43 − B cells were isolated using the B-cell Isolation Kit (Miltenyi; 130-090-862).

Techniques: Fluorescence, Staining