Journal: PLoS ONE
Article Title: Microglia in the Mouse Retina Alter the Structure and Function of Retinal Pigmented Epithelial Cells: A Potential Cellular Interaction Relevant to AMD
Figure Lengend Snippet: Immunoreactivity of cultured retinal microglia from CX3CR1 +/GFP transgenic mice. Retinal microglia isolated from CX3CR1 +/GFP transgenic mice exhibited GFP fluorescence (A) as driven by the microglial-specific CX3CR1 promoter. Cultured cells were immunoreactive for classic microglial markers Iba-1 (B), CD11b (C), F4/80 (D), and CD68(E). The cells were immunonegative for other macroglial markers such as GFAP (F), vimentin (G), and neuronal markers, such as NeuN (H). Scale bar = 100 µm.
Article Snippet: Cells or tissues were placed in a blocking solution (USB, Rodeo Blocker, Cleveland, OH) for 30 minutes, and incubated overnight with primary antibodies to the following antigens: CD11b, 1∶200; F4/80, 1∶200; CD68, 1∶200 (all from AbD Serotec, Raleigh, NC); ionized calcium binding adaptor molecule-1 (Iba1), 1∶500 (Wako, Richmond, VA); GFAP, 1∶200; Alexa Fluor 633-conjugated Phalloidin, 1∶100; zonula-occludins-1 (ZO-1), 1∶200 (all from Invitrogen, Carlsbad, CA); NeuN, 1∶200 (Millipore, Billerica, MA); vimentin, 1∶200 (MP Biomedicals, Solon, OH); CD31 [P2B1], 1∶100 (Development Studies Hybridoma Bank, Iowa City, IO).
Techniques: Cell Culture, Transgenic Assay, Mouse Assay, Isolation, Fluorescence