Figure 6.

Hyperglycemia upregulates C5aR expression on Müller cells. (A) MIO-M1 cells were cultured in high-glucose (30 mM) media for 7 days with daily media change, then cells were cultured in the absence or presence of 1 μM PGE₂ for another 24 hours, followed by flow cytometry analysis of C5aR expression on the surface of Müller cells. Dotted line, isotype control; solid line, anti-C5aR mAb staining. (B) MIO-M1 cells were cultured in normal (5 mM) or high-glucose (30 mM) media for 7 days with daily media change, then cultured in the absence or presence of 1 μM PGE₂ for another 24 hours. Then different amounts of C5a, with or without 50 μM C5aRA, were added into each well. Forty-eight hours later, IL-6 and VEGF levels in the culture supernatants were measured by conventional ELISA. *P < 0.05.