Specific pathogen free male c57bl 6 mice

Specific-pathogen-free male C57BL/6 mice (n=12, 8–10 weeks) were purchased from SPF Biotechnology Co. Ltd. (Beijing, China). All mice were housed in the Animal Experimental Center of Zhongnan Hospital at Wuhan University where the temperature was maintained at 25 °C with a 12 h light/dark cycle and free access to food and water. After adaptive feeding for 7 days, mice were randomly allocated to two groups (vehicle- and TMZ-treated groups; n=6/group) and underwent intracranial GL261-cell implantation and subsequent vehicle or TMZ gavage.
The mouse glioblastoma cell line GL261 was obtained from the American Type Culture Collection (Rockville, MD) and cultured in DMEM supplemented with 10% FBS in a humidified 5% CO₂ cell incubator at 37 °C.

Specific pathogen-free male C57BL/6 mice (n=36, 8–10 weeks old) were purchased from SPF Biotechnology (Beijing, China). All mice were housed in the Animal Experimental Center of Zhongnan Hospital at Wuhan University, where the temperature was maintained at 25°C with a 12 h light/dark cycle and free access to food and water. After adaptive feeding for 7 days, the mice were randomly allocated to four groups: control group (CON), TMZ group, LGG group, and LGG+TMZ group, with different treatments described below. Each group comprised nine mice.