https://orcid.org/0000-0001-8956-4918
Figures & data
Table 1. Differentially expressed miRNAs between RA cases and controls.
Figure 1. Networks constructed by the DEMIRs and DEGs.
(a). Regulatory networks constructed by all negatively correlated miRNAs and target mRNAs (listed in Table S1). (b). Regulatory networks constructed by highly correlated miRNA-mRNA pairs (r< −0.4).
Figure 2. Differential expressions of miRNAs in the validation sample.
The sample consisted of 35 RA cases and 35 healthy subjects. The expression level of each miRNA had been normalized against RNU48. P values were calculated by the two-sided Student’s t-test. * P < 0.05, ** P < 0.01. The miRNAs with non-significant differential expression or inconsistent direction with the discovery sample were shown in Figure S6.
Figure 3. Functional effect of miR-99b-5p overexpression on behaviours of Jurkat T cells.
(a). Relative expression of miR-99b-5p in OE cells and OE-NC cells using RT-qPCR. The expression was normalized against U6. (b). Cell proliferation using cell counting kit-8 assay. (c). Cell apoptosis using Annexin V/PI double staining. (d). Cell cycle using PI staining. (e). Cell activation assay with (+) or without (-) phytohemagglutinin (PHA) stimulation. (f). Expression changes of inflammatory cytokines. The expression level of each mRNA was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Two-sided Student’s t-test was used for the comparisons between groups. * P < 0.05, ** P < 0.01, *** P < 0.001. OE: over-expression cells, OE-NC: negative control cells.
Figure 4. Functional effect on behaviours of Jurkat T cells by using miR-99b-5p inhibitor.
(a). Relative expression of miR-99b-5p in miR-99b-5p inhibitor cells and inhibitor-NC cells using RT-qPCR. The expression was normalized against U6. (b). Cell proliferation using cell counting kit-8 assay. Two-sided Student’s t-test was used for the comparisons between groups. ** P < 0.01, *** P < 0.001.
Figure 5. Effect of miR-99b-5p on target gene expression.
(a). Relative mRNA levels of 7 target genes in miR-99b-5p OE cells and OE-NC cells using RT-qPCR. The expression was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). (b). Relative mRNA levels of 2 target genes in human PBMC in 35 RA cases and 35 controls using RT-qPCR. The expression was normalized against beta-2-microglobulin (B2M). (c). Dual luciferase assay of Jurkat T cells co-transfected with the luciferase reporter plasmid containing the wild-type or mutant-type RASSF4-3ʹUTR and miR-99b-5p overexpression lentivirus or negative vector. Two-sided Student’s t-test was used for the comparisons between groups. * P < 0.05, ** P < 0.01, *** P < 0.001. OE: over-expression cells, OE-NC: negative control cells.
