HepG2 cells (3×10
5 cells/well) were treated with extract (0.25 or 1 µg/mL), compounds or controls (0.5 or 2 µM) for 0, 4, 6 or 8 hours. After treatment, mRNA extraction was performed using the RNeasy Plus Mini Kit (Qiagen, Maryland, USA, Cat No. 74134). mRNA was quantified with a NanoDrop 2000c (Thermo Fisher Scientific, Waltham, MA). The expression of the
HMOX-1 and
NQO1 genes
21 (link),
39 (link)–
41 (link) was evaluated by RT-qPCR using predesigned TaqMan Gene Expression Assays (Hs01110250_m1 and Hs01045993_g1) and the TaqMan® RNA-to-CT
TM 1-Step Kit (Applied Biosystems, Foster City, CA, Cat No. 4331182). The run method was holding at 48°C for 15 min, 95°C for 10 min and cycling (40 cycles) at 95°C for 15 sec and 60°C for 1 min. β-actin (Applied Biosystems, Foster City, CA, ref. 4325788) was used as an endogenous control.
5 cells/well) were treated with extract (0.25 or 1 µg/mL), compounds or controls (0.5 or 2 µM) for 0, 4, 6 or 8 hours. After treatment, mRNA extraction was performed using the RNeasy Plus Mini Kit (Qiagen, Maryland, USA, Cat No. 74134). mRNA was quantified with a NanoDrop 2000c (Thermo Fisher Scientific, Waltham, MA). The expression of the
HMOX-1 and
NQO1 genes
21 (link),
39 (link)–
41 (link) was evaluated by RT-qPCR using predesigned TaqMan Gene Expression Assays (Hs01110250_m1 and Hs01045993_g1) and the TaqMan® RNA-to-CT
TM 1-Step Kit (Applied Biosystems, Foster City, CA, Cat No. 4331182). The run method was holding at 48°C for 15 min, 95°C for 10 min and cycling (40 cycles) at 95°C for 15 sec and 60°C for 1 min. β-actin (Applied Biosystems, Foster City, CA, ref. 4325788) was used as an endogenous control.
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