Trizol reagent 15596026

Bisphenol A [2,2-bis (4-hydroxyphenyl) propane] (BPA) (239658) and nonylphenol (NP) (442873) were obtained from Sigma (St. Louis, MO, USA). LipofectAMINE 2000 (11668-027), p-nitrophenyl phosphate (p-NPP) (002212), substrate of alkaline phosphatase, TRIzol-Reagent (15596026), FURA2-AM (F1221), and propidium iodide (P1304MP) were obtained from Invitrogen (Carlsbad, CA, USA). BB-94 (Batimastat) (196440) was obtained from Calbiochem (San Diego, CA, USA). ADAM17 antibody (ab39163) was purchased from Abcam (Cambridge, MA, USA). PARP-1/2 antibody (sc-7150) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cleaved Caspase-3 (Asp175) antibody (#9661) was acquired from Cell Signaling (Danvers, MA, USA). β-actin (AC-15) and anti-rabbit IgG-FITC (F0382) antibodies were purchased from Sigma (St. Louis, MO, USA). Peroxidase anti-mouse IgG (5220-0286) and peroxidase anti-rabbit IgG (5220-0283) antibodies were obtained from KPL (Gaithersburg, MD, USA). Human phospho-kinase antibody array (Catalog #ARY003B) was obtained from R&D Systems (Minneapolis, MN, USA). Western Lightning Chemiluminescense Reagent Plus kit (NEL104001EA) was obtained from PerkinElmer Inc. (Waltham, MA, USA).

The RNA from cells, plasma, and brain tissue was extracted by TRIzol kit procedure (Invitrogen™ TRIzol™ Reagent 15596026), and cDNA was synthesized by reverse transcription (Invitrogen™ M-MLV Reverse Transcriptase 28025013) after passing the quality inspection, and the PCR reaction system was configured to detect the mRNA expression of lncRNA GAS5. The primer sequences are lncRNA GAS5 forward-5′-TCT AGC TTG GGT GAG GCA-3′ and reverse-5′-TGG AGA GTC GGC TTG ACT A-3′;GAPDH forward-5′-ACG GAT TTG GTC GTA TTG G-3′ and reverse-5′-TCC CGT TCT CAG CCT TG-3 ′; lncRNA GAS5 forward-5′-TCT AGC TTG GGT GAG GCA-3′ and reverse-5′-TGG AGA GTC GGC TTG ACT A-3′; and GAPDH forward-5′-ACG GAT TTG GTC GTA TTG G-3′ and reverse-5′-TCC CGT TCT CAG CCT TG-3′. For 50 μL total volume, the following PCR recipe was used: 10x buffer 4 μL, 50 mM MgCl₂ 2.5 μL, amplification primer 1 (10 μM) 0.9 μL, amplification primer 2 (10 μM) 0.9 μL, Taq DNA polymerase (5 U/μL) 0.5 μL, cDNA 2.2 μL, and dis.H₂O 39 μL. For the PCR profile, initial denaturation was done at 94°C, denaturation at 96°C, annealing at 56°C, elongation at 71°C, and final elongation at 72°C.