Fig. 6
A A cluster heatmap showing methylation changes in promoter of 13 microRNAs between LGI1 encephalitis cohort and normal donors. B Morphological characteristics of apoptotic body, microvesicles, and exosome isolated from serum of one LGI1 encephalitis patient by TEM. C Particle size of microvesicles and exosome using NTA. D Protein markers of apoptotic body (C3B, C1QC), microvesicles (ARF6), exosome (CD63, TSG101) and serum supernatant by western blot. E Volcano plot of differentially expressive exosome-microRNAs between LGI1 encephalitis and healthy control. F A scatter plot assessing the expression variation of exosome microRNAs between LGI1 encephalitis patients and healthy donors. G Venn diagram showing the overlap of 71 differential exosome microRNAs and 13 methylated-driven microRNAs. H The methylation changes in promoter of hsa-miR-2467-5p between LGI1 encephalitis cases and healthy donors. I PCR analysis of miR-2467-5p expression in PBMCs between LGI1 encephalitis patients and healthy donors. J PCR analysis of miR-2467-5p expression in apoptotic bodies isolated from LGI1 encephalitis patients and healthy donors. K PCR analysis of miR-2467-5p in microvesicles isolated from LGI1 encephalitis patients and healthy donors. L PCR analysis of miR-2467-5p in exosomes isolated from LGI1 encephalitis patients and healthy donors. M Schematic representation of the complementary binding sites of CSF3 and PDCD1 with miR-2467-5p. Relative luciferase activity of wild-type and 3`UTR mutant constructs of CSF3 N and PDCD1 O cotransfected with miR-2467-5p mimics and miRNA-NC. PCR analysis of PDCD1 P, CSF3 Q and CCL15 R expression in PBMCs after transfection of miR-2467-5p mimics, inhibitor or scramble control into PBMCs. ELISA analysis of PDCD1 S, CSF3 T, and CCL15 U expression in cell supernatants after transfection of miR-2467-5p mimics, inhibitor or scramble control into PBMCs. Scatterplots showing the association of miR-2467-5p in PBMCs with the expression of PDCD1 V, CSF3 W, and CCL15 X in serum in LGI1 encephalitis patients (n = 6) and healthy donors (n = 4). LGI1-E LGI1 encephalitis; ND normal donor; AE autoimmune encephalitis; MV microvesicles; ABs apoptotic bodies; Sup supernatant; NC negative control. Data are means ± SD of three experiments. *p < 0.05, **p < 0.01, ***p < 0.001