Figure 5

CRISPR/Cas9-mediated disruption of CD38 does not phenotypically affect RA-induced differentiation. (A) Wild-type and CRISPR-derived cell lines were cultured for 72 h with 1 μM RA as indicated and membrane CD38 expression was evaluated by flow cytometry at 24, 48, and 72 h. Gating to discriminate positive cells was set to exclude 95% of untreated wild-type cells. (B) Membrane CD11b expression was analysed at 24, 48, and 72 h using flow cytometry. *p < 0.05; **p < 0.01. Two-way ANOVA using Tukey’s multiple comparisons test was used to determine significance. (C) Cell density was measured at 24, 48, and 72 h using a haemocytometer and 0.2% Trypan Blue exclusion staining. (D–F): Cell cycle distribution was analysed using flow cytometry with propidium iodide staining at 24, 48, and 72 h. Error bars indicate SEM.