Human normal astrocyte line SVGp12 and human GBM cell lines U87 and U118 (Cell Bank of Chinese Academy of Sciences, Shanghai, China), and LGG cell line HS683 (Procell, Wuhan, China) were cultured in DMEM medium (Biowest, FRA) containing 10% fetal bovine serum (HyClone, USA) at 37 °C and 5% CO2.
Total RNA was extracted from SVGp12, HS683, U87, and U118 cells using TRIzol (Invitrogen, USA). cDNA was synthesized using the PrimeScript™ RT Reagent kit (Takala, Japan). cDNA was extracted from SVGp12, HS683, U87 and U118 cells using THUNDERBIRD® SYBR® qPCR Mix (TOYOBO, Japan) for real-time PCR to detect TASL expression, and the experiments were repeated at least three times and calculated by the 2−△△Ct method. The reaction conditions were as follows: 95 ℃ preheat for 5 min; 95 ℃ for 15 s, 57 ℃ for 30 s, 72 ℃ for 30 s, 5 cycles; 95 ℃ for 15 s, 59 ℃ for 30 s, 72 ℃ for 30 s, 35 cycles.
The following primers were used:
Total RNA was extracted from SVGp12, HS683, U87, and U118 cells using TRIzol (Invitrogen, USA). cDNA was synthesized using the PrimeScript™ RT Reagent kit (Takala, Japan). cDNA was extracted from SVGp12, HS683, U87 and U118 cells using THUNDERBIRD® SYBR® qPCR Mix (TOYOBO, Japan) for real-time PCR to detect TASL expression, and the experiments were repeated at least three times and calculated by the 2−△△Ct method. The reaction conditions were as follows: 95 ℃ preheat for 5 min; 95 ℃ for 15 s, 57 ℃ for 30 s, 72 ℃ for 30 s, 5 cycles; 95 ℃ for 15 s, 59 ℃ for 30 s, 72 ℃ for 30 s, 35 cycles.
The following primers were used:
TASL forward (5'-ACAAGTCAGAAGTCTCTACGTG-3');
TASL reverse (5'-TCTCTCTGACTTCTGCTATGTTGG-3');
GAPDH forward (5'-CCTGACCTGCCGTCTAGAAA-3');
GAPDH reverse (5'-AGGAAAAGCAG-GAGGGTAGC-3').