Dmem high glucose medium

Toxoplasma gondii tachyzoites of both the RH and ME49 strains were cultured in DMEM/high-glucose medium (Servicebio, Wuhan, China) containing 10% fetal bovine serum (FBS) and incubated with Vero cells in an environment of 37 °C with 5% CO₂.

HepG2 and SMMC-7721 cells were purchased from ATCC. HepG2 cells were cultured in DMEM high glucose medium (Servicebio), and SMMC-7721 cells were cultured in 1640 medium (Servicebio), all containing 10% Foetal Bovine Serum (FBS) and 1% penicillin and streptomycin (Servicebio). Short hairpin (sh)RNAs sh-STK25 were obtained from miaolingbio.lnc, Wuhan, China. HepG2 and SMMC-7721 cells were cultured in 6-well plates (5 × 10⁵/well) and transfected with 2.4 μg siRNA using Attractence Transfection Reagent (Cat. No. 301005, QIAGEN, China).

Artemisinin was provided by Solarbio (Beijing, China). DMEM/high-glucose medium was obtained from Servicebio (Wuhan, China). The following primary antibodies were used: NF-κB p-65 (1:1000) and phosphorylated NF-κB p-65 (1:700) were procured from ABclonal (Wuhan, China); p-38 (1:1000), phosphorylated p38 (1:700), IKK (1:1000), and phosphorylated IKK (1:700) were purchased from CST (Boston, MA, USA); and TLR4 (1:1000), Myd88 (1:1000), β-actin (1:1000), GAPDH (1:80000), and Tubulin (1:50000) were purchased from Proteintech (Wuhan, China). The secondary antibodies, HRP-conjugated Affinipure Goat Anti-Rabbit IgG (1:5000) and HRP-conjugated Affinipure Goat Anti-Mouse IgG (1:5000), were obtained by Proteintech (Wuhan, China). The RT-qPCR-related reagents were provided by Accurate Biotechnology (Hunan, China).