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. 2021 Mar 8;12:633115. doi: 10.3389/fneur.2021.633115

Figure 1.

Figure 1

Cultured human cells (HEK293 cells) were transfected with the antigen, MOG or AQP4, and used as substrate in an indirect immunofluorescence assay. For MOG-IgG detection, cells were stably transfected with hMOG-α1 splice variant complementary DNA (cDNA) (FL-MOG-α1) cloned into pCMV6-AC-GFP plasmid. (B) Cells were stably transfected with hMOG construct or empty vector and used as the substrate for live cell-based assays (CBAs). (A) HEK-293 cells were also transfected with pmCherry-hAQP4-M23 expression vector, cloned to obtain M23mCherry stable expressing cell line. (B) Serum MOG-IgG was detected on the surface of MOG expressing cells, using goat antihuman 568 Alexa-Fluor-conjugated secondary antibodies, (A) while AQP4-IgG bound antibodies were detected using goat antihuman 488 Alexa-Fluor-conjugated secondary antibodies. AQP4-M23-IgG and MOG-IgG binding was determined using live-cell-staining immunofluorescence technique. MOG, myelin oligodendrocyte glycoprotein; AQP4, aquaporin-4 channel protein; HEK-293, human embryonic kidney 293 cells.