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. 2022 Dec 6;15:6607–6616. doi: 10.2147/JIR.S383623

Figure 3.

Figure 3

miR-31-5p directly targets SLC15A4 and negatively regulates the IRF5 pathway. (A) Sequence alignment of miR-31-5p and its putative binding sites in the 3′UTR of SLC15A4. Mutations of the miR-31-5p target sites are indicated on top. (B) Luciferase activity in HEK-293 cells transfected with SLC15A4-WT or SLC15A4-MT, cotransfected with mimics control (NC-mimic) or miR-31-5p-mimics (miR-31-5p-mimic). Error bars represent the SEM. Student’s t-test, ns p > 0.05 and ***p < 0.001. (C) THP-1 and CAL-1 cells were transfected with negative controls (NC-mi and NC-in), miR-31-5p mimics (miR-31-mi) or miR-31-5p inhibitor (miR-31-in), respectively, and then stimulated with R848 (5µg/mL) for 24h. Whole-cell lysates were prepared, and Western blot was performed for TASL, IRF5 and GAPDH. (D) CAL-1 cells were transfected with negative controls (NC-mimics and NC-inhibitor, NM and NI), miR-31-5p mimics (31-mi) or miR-31-5p inhibitor (31-in) and stimulated with R848 (5µg/mL) for 2h and 4h respectively. Whole-cell lysates were prepared and Western blot was performed for p-IRF5 and GAPDH.