Analysis of CD11b differentiation marker by flow cytometry.
(A) HL-60 cells were plated at 1×105 cells/mL and differentiation induced. Undifferentiated cells received vehicle solution for the retinoic acid. Differentiated and undifferentiated cells were collected at the indicated time points and processed for flow cytometric analysis of CD11b positive cells. * P<0.05 compared with undifferentiated time-matched samples. (B) Effect of modulation of either SMS1 or SMS2 on differentiation of HL-60 cells by analysis of CD11b differentiation marker using flow cytometry. Two million HL-60 cells were transfected with 4.5 µg of SCR siRNA, SMS1 siRNA (1.2 and 1.4), and SMS2 siRNA (2.0, 2.3) by nucleofection. Downregulation and differentiation proceeded for 48 hours. Cells were then collected and processed for flow cytometric analysis of CD11b. * P<0.05 compared with SCR siRNA undifferentiated cells. UD: undifferentiated cells; D: differentiated cells; CT MOCK: control for transfection reagent; SCR: control scrambled siRNA.
