Journal: Open Medicine

Article Title: EphA3 targeted by miR-3666 contributes to melanoma malignancy via activating ERK1/2 and p38 MAPK pathways

doi: 10.1515/med-2022-0597

Figure Lengend Snippet: Real-time PCR primer synthesis list

Article Snippet: Proteins (20 μg per lane) were resolved in 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and detected on poly(vinylidene fluoride) membranes after which was blocked by 5% skim milk powder at room temperature for 2 h. Protein detection on membranes was detected with anti-p38MAPK (FNab06077, 1:1,000, FineTest, Biological Technology Co., Ltd, China), anti-p-p38MAPK (44-684G, 1:1,000, ThermoFisher), anti-p-ERK1/2 (44-680G, 1:1,000, ThermoFisher), anti-ERK1/2 (13-6200, 1:1,000,ThermoFisher), anti-EphA3 (310438-T08, 1:1,000, SinoBiological, China), and anti-GAPDH (100242-T08, 1:1,000, SinoBiological) at 4°C for 24 h. Next, horseradish peroxidase-conjugated secondary antibodies (A32731, 1:1,000, ThermoFisher) were added and incubated with these membranes at room temperature for 1 h. Immune blots were developed with enhanced chemiluminescence (GE Healthcare, USA) and counted quantitatively by ImageJ.

Techniques: Real-time Polymerase Chain Reaction