She designe d the dolls as her service project for the school's National Honor Society chapter. The four doll recipients (接受者) are from B. She designe d an d made dolls for them.Fresh Start Surgical Gifts in Carlsbad. The 19-year-ol d organization provides C. She gave them some ...
Cells were transfected with siRNAs using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. siRNAs targeting CD147 were designed and synthesized by Shanghai GenePharma Co. (Shanghai, China). The siRNA sequence is depicted in Table S1. GSK-3β siRNA...
Real-time PCR was carried out on the StepOnePlus Real-time PCR System (Applied Biosystems, Life Technologies, Carlsbad, CA, USA) with SYBR Premix Ex Taq (TaKaRa). The sequences of primers were: miR-200b forward: 5′-TGCCGTAATACTGCCTGGTAA-3′, reverse: 5′-CAGAGCAGGGTCCGAGGTA-3′;...
Total RNA was extracted with TRIzol reagent (Invitrogen, Carlsbad, CA, USA), and the overall quality was assayed by agarose gel electrophoresis. The cDNA was obtained using a PrimeScript™ RT Reagent Kit with gDNA Eraser (Takara). Next, real-time PCR was conducted with Applied Biosystems Quan...
PCR was performed using Amplitaq Gold polymerase (Applied Biosystems, Carlsbad, CA), with a PCR cycle of 94 °C for 10 min followed by 35 cycles of 94 °C for 15 s, 60 or 65 °C for 30 s, and 72 °C for 30 s, followed by 7 min at 72 °C. Products were visualized with ...
Sample integrity was verified using the RNA Pico Assay (Life Technologies, Carlsbad, CA) and Nano Drop (Thermo-Fisher), while the sample concentration was determined with the Quant-It RNA Assay (Life Technologies). Removal of DNA was confirmed using the Quant-iT High-Sensitivity DNA Assay (...
Total RNA was extracted using TRIzol reagent and resuspended in RNase-free water (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. Samples with an A260/280 ratio ≥ 1.8 were selected and stored at − 70 °C until use. 2.5 Microarray Analysis RNA integri...
(Invitrogen, Carlsbad, CA) according to the manufacturer's instructions.Transfected cells were treated with 0.5 and 1 渭mol/L PAB for indicated times.Western Blot Analysis: Expression of proteins were detected by western blot method.Cell lysate from various experiments containing 10-80 mg protein ...
coli Omnimax (Invitrogen, Carlsbad, CA, USA) in 2YT (16 g Bacto tryptone, 10 g Bacto yeast extract and 5 g NaCl, per liter water) to each well and incubation for 30 min at 37 °C with shaking. M13K07 helper phage (New England Biolabs (NEB), Ipswich, MA, USA) was ...
All cells were transfected using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. siRNAs, miRNA mimics and mutant miRNAs were synthesized by GenePharma (Suzhou, China) and miRNA inhibitors purchased from RiboBio (Guangzhou, China). The siRNA ...