Automatic enzyme label analyzer

To evaluate the proliferation of BV-2 cells, the cells were counted by Cell Counting Kit (CCK-8, Dojindo, Japan). Briefly, the cells were seeded into 96-well plates at a density of 1 × 10⁴ cells per well in 100 μl medium; and then cells were transfected with shRNA and overexpression plasmid by Micropoly-transfecter^TM Cell Reagent. Subsequently, 10 μl of CCK-8 reagent was added to each well and incubated at 37°C for 1–2 h. The absorbance of optical density was determined using the Automatic Enzyme Label Analyzer (Allsheng, China) at 450 nm (A450). Each experiment was performed in triplicate and repeated three times to ensure accuracy.

To evaluate the proliferation of BV-2 cells, the cells were counted by Cell Counting Kit (CCK-8, Dojindo, Japan). The 96-well plate was seeded with 100 μl of BV-2 cell suspension, and was incubated in a 5% CO₂ incubator at 37°C. When the cells grew to 70–80%, cells were transfected with interference and overexpression plasmids by Micropoly-transfecter™ Cell Reagent. After 12, 24, 36, 48, and 60 h of incubation, 10 μl of CCK-8 reagent and 100 μl of fresh DMEM cell medium were added to each well and incubated at 37°C for 1–2 h. Subsequently, the absorbance value was detected at 450 nm by the Automatic Enzyme Label Analyzer (Allsheng, China). Each experiment was performed in triplicate and repeated three times to ensure accuracy.