Dual Functions of the C5a Receptor as a Connector for the K562 Erythroblast-Like Cell-THP-1 Macrophage-Like Cell Island and as a Sensor for the Differentiation of the K562 Erythroblast-Like Cell during Haemin-Induced Erythropoiesis

Figure 4

Contribution of the C5aR-RP S19 oligomer system to the haemin-induced terminal differentiation of K562 cells under monocell culture conditions. K562 cells expressing EGFP-ANXA3/Mock, EGFP-ANXA3/Wild-type RP S19, EGFP-ANXA3/Q137N mutant RP S19, and EGFP-ANXA3/RGS3 were harvested at several time points after the haemin induction and analysed under CLSM (𝑛=3). The mock (black), Wild-type RP S19 (red), Q137N mutant RP S19 (green), and RGS3 (yellow) K562 cells were harvested at several time points after the haemin induction and stained with FITC-conjugated anti-CD71 and APC-conjugated anti-CD235a mouse IgGs for FACS analyses (𝑛=3). The averages of the percentage and number of CD235a⁺/CD71⁻ erythrocyte-like cells were calculated on day 20 of the monocell culturing. 𝑃 values of less than 0.05 were considered statistically significant (*𝑃<0.05 or **𝑃<0.01).