FIG 2.

Knockdown of glycophorins A, B, and C in cultured erythrocytes. (A) Expression of glycophorin A (GPA), glycophorin B (GPB), and glycophorin C (GPC) on the cell surface of GPA knockdown (KD) (green), GPB KD (blue), and GPC KD (pink) cultured erythrocytes (cRBCs) as determined by flow cytometry. Representative flow cytometry plots are shown. Gray traces, receptor expression on pLKO cRBCs; dashed traces, pLKO cells stained with a control antibody. (B) Mean expression ± standard error from three to seven independent experiments, normalized to expression on pLKO cells. For measurement of GPB expression, cRBCs were treated with trypsin to remove GPA and cells stained with a GPA/B antibody. Statistical significance was determined using a one-way analysis of variance (ANOVA) with a Dunnett multiple-comparison test. ***, P ≤ 0.001. (C) May-Grünwald- and Giemsa-stained cytospins showing normal morphology of KD and pLKO control cRBCs. cRBCs were passed through a 5-μm filter to remove nucleated cells prior to flow cytometry and cytospin preparation.