Fig. 6: Neat1 promotes antigen-specific Th17 cell responses by regulating miR-128-3p/NFAT5 axis.

A Venn diagram analysis of putative Neat1 target miRNAs using Starbase v3.0, DIANA-LncBase, and miRDB databases. B Only 23 candidate miRNAs were expressed in CD4⁺ T cells isolated from EAU mice. Microarray-based bar graphs showing the expression level of candidate miRNA in CD4⁺ T cells isolated from EAU mice relative to those from naïve mice. C Real-time qRT-PCR analysis of miR-128-3p expression in Th17 cells transfected with ASO-NC or ASO-Neat1 (n = 4 per group). D Sequence alignment between miR-128-3p and its potential binding sites (in red letters) in the lncRNA Neat1. E, F Luciferase activity analysis of reporter carrying Neat1 binding sites or mutant site (Mut) co-transfected into HEK293T with Ctrl mimics or miR-128-3p mimics (n = 4 per group). G–K CD4⁺ T cells isolated from immunized mice were transfected with indicated oligonucleotides and stimulated with IRBP_1-20 in the presence of irradiated APCs under Th17-polarizing conditions. G, J Real-time qRT-PCR analysis of Rorc, Il17 and Il23r expression (n = 4 per group). H ELISA analysis of IL-17 secretion in the culture supernatants (n = 4 per group). I, K Real-time qRT-PCR analysis of Nfat5 expression (n = 4 per group). Data are presented as mean ± SD of at least three independent experiments. *p < 0.05, **p < 0.01. See also Fig. S3.