Figure 2

NF54 GIE fail to adhere to erythroid cells. (a) Cell-cell adhesion assays of MACS-purified NF54 GIE (GIE) or uninfected erythrocytes (uE) to human primary erythroblasts, UT7 cells and K562 cells. Error bars denote the standard error of the mean. ns, non-significant differences in adhesion rates. n = number of experiments. (b) Live microscopy imaging showing non-specific adhesion of MACS-purified NF54 GIE (upper panel) and uninfected erythrocytes (lower panel) stained with PKH67 (green) to human primary erythroblasts or K562 cells stained with PKH26 (red). DNA is stained with Hoechst 33342 (blue). Bars represent 5 µm. (c) Immunofluorescence analysis (IFA) of paraformaldehyde-fixed uninfected erythrocytes, erythroblasts, K562 cells and UT7 cells stained with anti-GPC mAb followed by anti-mouse Alexa594-conjugated IgG, showing the presence of GPC at the surface of all erythroid cells. DNA is stained with Hoechst 33342 (blue). Bars represent 5 µm. (d) Western blot analysis showing the presence and the size of GPC in erythrocytes, erythroblasts, K562 cells and UT7 cells. Red star indicates the expected size of fully glycosylated GPC (32 kDa). Black stars indicate degradation products. Full-length blot is presented in Supplementary Fig. S1.