Penicillin streptomycin

Mouse microglia BV2 cells were grown in high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM) (Biological Industries, Iseral) supplemented with 10% fetal bovine serum (FBS) (Lanzhou Bailing Biotechnology, China) and penicillin/streptomycin (BBI Life Sciences, China). The cells were maintained in a humidified incubator (Sanyo, Japan) with a 95% air/5% CO₂ atmosphere at 37 ^°C. (+)-JQ1 (Nanjing Tianzhun, China) was dissolved in DMSO to a final concentration ranging from 0 to 10 μM. In the experiments indicated, the cells were pretreated with various concentrations of either (+)-JQ1 or DMSO 30 min prior to LPS (100 ng/ml) (Sigma-Alorich, USA)/Aβ (10 μM) (Sigma-Alorich, USA) treatment. Minomycine (2 μM) (Shyuanye, China) was administrated as the positive control. For NFκB signaling test, PDTC (p65 inhibitor), U0126 (ERK inhibitor) or SB203580 (p38 inhibitor) (Calbiochem, Germany) were treated with LPS or (+)-JQ1 for 24 h, followed by medium collection.

RK13 rabbit epithelial cells (American Type Culture Collection CCL-37) were transfected with mammalian expression vector pcDNA3.1(+)-moPrPWT to express the full-length PrP^C. A separate set of RK13 cells was transfected with pcDNA3.1(+)-moMet4-1 or pcDNA3.1(+)-moΔN6 to express mutant PrP Met4-1 and ΔN6, respectively. These expression vectors were constructed by Generay Biotechnology. All plasmids were linearized by digestion with ScaI (Thermo Fisher Scientific; catalog number: FD0434) before transfection. All transfected cells were selected with 600 to 1200 μg/ml of Geneticin (Gibco; catalog number: 11811023) in Dulbecco's modified Eagle's medium (Gibco; catalog number: 11965092) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (Bovogen; catalog number: SFBS), 1% (v/v) l-glutamine (BBI Life Sciences), and 1% (v/v) penicillin/streptomycin (BBI Life Sciences; catalog number: E607011) as described previously (54 (link)).