Mtt cell proliferation and cytotoxicity assay kit

The MTT Cell Proliferation and Cytotoxicity Assay Kit (Dojindo Laboratories, Tokyo, Japan) were used to assess the proliferation activity of human glioma cell lines (LN229 and U251) according to the manufacturer’s instructions. Cell lines were all culture at 96-well plates with a density of 1×10⁴ cells/well and then transfected with siRHPN1-AS1, miR-625-5p mimics, or miR-625-5p inhibitor. The cells were incubated for 12, 24, 48 and 72 hrs. After incubation, the MTT solution was removed and replaced with dimethyl sulfoxide (DMSO; 150 μL, 4%; Sigma). A microplate reader (Bio-Tek, Instruments, Neufahrn, Germany) was used to measure the absorbance at 490 nm.

The cell viability of human glioma cell lines (U138 and U251) was detected using the MTT Cell Proliferation and Cytotoxicity Assay Kit that purchased from Dojindo Laboratories (Tokyo, Japan) according to the manufacturer’s instructions. Cell lines (1×10⁴ cells/well) were all cultured in 24-well plates and then transfected with Vector/NR5A2 or Scramble/NR5A2-sh2#. The transfected U138 and U251 cells were cultured for 12, 24, 48, and 72 h. After that, the MTT solution was removed and replaced with 150 μL 4% dimethyl sulfoxide (DMSO; Sigma). Moreover, the half maximal inhibitory concentration (IC₅₀) was detected to determine the cytotoxicity of TMZ. A microplate reader (Bio-Tek, Instruments, Neufahrn, Germany) was used to measure the absorbance at 490 nm. Experiments were done in triplicate.