Temozolomide tmz

The cell lines U251 and U87 from humans were procured from the ATCC (Manassas, VA) and grown in RPMI 1640 medium along with foetal bovine serum (5%) in humid air with 5% CO₂ at 37℃. Stable Mcl‐1 overexpression was achieved by infecting a given cell line for 24 hours with lentiviruses carrying empty vector or Mcl‐1 as described previously.¹⁷ (link), ¹⁸ (link) On receiving the cells, annual detection of mycoplasma was performed using the Mycoplasma Detection Kit Nozan MycoAlert (Sigma, USA) to make sure that they were free from any mycoplasma. SD‐36 and Temozolomide (TMZ) were obtained from MedChemExpress. S63845 was purchased from Chemietek (Indianapolis, IN).

In total, 25 eight-week-old female BALB/c AnN.Cg-Foxnlnu/CrlNarl mice were purchased from the National Laboratory Animal Center, Taipei, Taiwan. One week later, 1 × 10⁵ GBM8401-Luc tumor cells were implanted into the right cerebral hemisphere of the mice. The animals were randomly divided into four groups: shLuc, shLuc + Temozolomide (TMZ, MedChem Express, NJ, USA), shKDELC2, and shKDELC2 + TMZ treatment groups. TMZ was administered through oral gavage at 5 mg/kg/day for 7 days. The region of interest (ROI) was monitored using a noninvasive in vivo imaging system (IVIS) (Perkin Elmer, Massachusetts, USA) and measured at 0, 2, 5, and 7 days of TMZ administration. The bioluminescence intensity was compared after intraperitoneal injection of D-luciferin firefly and potassium salt (Biosynth, Thal, Swiss) in PBS. At 7 days, the mice were euthanized with tiletamine-zolazepam and xylazine, and the brains were fixed in 10% formalin, embedded in paraffin, and cut into serial sections. Animal experiments were approved by the Institutional Animal Care and Use Committee of the National Defense Medical Center, Taiwan (Approval number: IACUC-20-106, Date: 14 April 2020).