Superpico ecl chemiluminescence kit

As described previously, the protein of systemic leaves was extracted from PVX-infected plants (Xiong et al., 2009 (link)). Immunoblotting was performed with primary Mouse anti-PVX CP monoclonal antibodies, followed by HRP-conjugated Goat Anti-Mouse IgG (Cat#: D110087, Sangon Biotech, Shanghai, China). Blotted membranes were washed thoroughly and visualized using the SuperPico ECL Chemiluminescence Kit according to the manufacturer’s protocol (Cat#: E422; Vazyme, Nanjing, China).

Total cellular proteins were extracted using RIPA lysis buffer (Beyotime), separated via electrophoresis on a 10% SDS separation gel (YEASEN, China), and transferred to PVDF membranes (Millipore, Germany). The PVDF membranes were then incubated with primary antibodies, including anti-SRSF5 (1:1000; Abcam, UK), anti-HOXD8 (1:1000; Santa Cruz, USA), anti-TRIM72 (1:2000; Proteintech, USA), anti-PKM2 (1:1000; CST, USA), and anti-GAPDH (1:5000; ABclonal, China) antibodies at 4 °C. Subsequently, the membranes were incubated with Peroxidase AffiniPure Goat Anti-Rabbit/Mouse IgG (H + L) (1:10000; Jackson ImmunoResearch, USA) for 1 h at room temperature. The protein bands were visualized with a SuperPico ECL Chemiluminescence Kit (Vazyme) and imaged using a ChemiDoc XRS + Imaging System (Bio-Rad, USA).