Mir 183 5p mimic

Transfections were performed using Lipofectamine RNAiMAX (Invitrogen Corporation, Carlsbad, CA, USA) according to the manufacturer's instructions. Short interfering (siRNA) targeting lncRNA-ERGAL and miR-183-5p inhibitors were transfected into HUVECs at a final concentration of 100 μM, whereas themiR-183-5p mimic was introduced at a final concentration of 50 μM. si-lncRNA, si-NC, miR-183-5p mimic, miR-183-5p inhibitor, and miR-NC were all designed and synthesized by RiboBio (RiboBio Co., Guangzhou, China) (Supplementary Table 3). Cells were then infected with DENV at 24 h after transfection.

miR-96-5p mimic, miR-183-5p mimic, and miR-142a-5p inhibitor, as well as their corresponding negative control (NCs), were designed by RiboBio Co., Ltd. (Guangzhou, China). BV-2 microglial cells (1.5×10⁵ cells/ml) in a 6-well plate were transfected with 50 nM miR-96-5p mimic, 50 nM miR-183-5p mimic, 100 nM miR-142a-5p inhibitor or their NCs by using a Lipofectamine 3,000 reagent (Invitrogen, Carlsbad, CA, United States) according to the manufacturer’s instructions. Transfected cells were incubated for an additional 24 h prior to the scratch injury. Corresponding sequences were as follows: miR-96-5p mimic, 5′UUU​GGC​ACU​AGC​ACA​UUU​UUG​CU3’; miR-183-5p mimic, 5′ UAU​GGC​ACU​GGU​AGA​AUU​CAC​U 3’; mimic-NC, 5′ UUU​GUA​CUA​CAC​AAA​AGU​ACU​G3’; miR-142a-5p inhibitor, 5′ GUA​UUU​CAU​CUU​UCG​UGA​UGA 3’; inhibitor-NC, 5′ CAG​UAC​UUU​UGU​GUA​GUA​CAA​A 3’. The efficiency of transfections was validated by comparing the levels of miR-96-5p, miR-183-5p, and miR-142a-5p between transfected and controlled cells by quantitative real-time-polymerase chain reaction (qRT-PCR).

The NR8383 cells and BMDMs were plated in a 6-well plate. When cells reached 50% confluence, they were transfected with 10 nM NC-mimic, miR-183-5p mimic, NC-inhibitor, miR-183-5p inhibitor, siFoxO1 or si-NC (RiboBio, Guangzhou, China). Lipofectamine 3000 (11668019, Thermo) was used for transfection experiments according to the product instructions.