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. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Exp Eye Res. 2018 Feb 16;170:29–39. doi: 10.1016/j.exer.2018.02.014

Fig 6.

Fig 6

CD40L-induced MCP-1 mRNA synthesis and protein secretion by hRPE cells. MCP-1 mRNA levels measured by RT-PCR in hRPE cells treated with or without IFN-γ pre-treatment (Pre-IFN) or control (Ctrl) for 24 hr and then switched to serum-free media containing 0 or 5 µg/ml of CD40L for 6 hr (A). The fold changes were calculated by normalization against β-actin and comparison with untreated control. MCP-1 ELISA of conditioned media from hRPE cells pre-treated with or without IL-1β (20 pg/ml) for 24 hr, then replaced with media with or without CD40L for another 24 hr (B). MCP-1 ELISA of conditioned media from hRPE cells treated with or without CD40L (5 µg/ml) in the presence or absence of anti-IL-1β, anti-CD11b, anti-α5β1 or corresponding isotype control antibody for 24 hr (C). MCP-1 ELISA of conditioned media from hRPE cells treated with 10% serum media containing 0 or 5 µg/ml of CD40L in the presence or absence of anti-CD40 or corresponding isotype control antibody for 24, 48 and 72 hr (D). MCP-1 ELISA of conditioned media from hRPE cells treated with 10% serum media with or without IFN-γ for 24 hr, then replaced with CD40L containing or CD40L-free media in the presence or absence of anti CD40, α5β1 or CD11b antibody or isotype controls for an additional 24 hr (E). The concentration of IFN-γ (IFN) was 500 U/ml. *p<0.05; **p<0.01; ***p<0.001, as compared with untreated control (CD40L-treated in C, pre-IL-1 or CD40L-treated in D, and pre-IFN or CD40L-treated in E) or between presence and absence of antibodies (B, CD40Ab in E).