Senp3

The following antibodies were used in this study: mouse monoclonal antibodies against β-actin (Sigma); Rabbit polyclonal antibodies against Sp1 (Santa Cruz Biotechnology); mouse monoclonal [Ubi-1] antibodies against Ubiquitin (Abcam), the rabbit polyclonal antibodies against HA (Abcam), the mouse monoclonal antibodies against GFP (Abcam), mouse anti-RH (Qiagen), the mouse monoclonal antibodies against FLAG (sigma), the rabbit monoclonal antibodies against SUMO2/3 (Cell Signaling Technology), normal rabbit IgG (CST), the rabbit polyclonal antibodies against RNF4 (Abgent), and the rabbit polyclonal antibodies against SENP3 (Protein Techgroup).

BCA Protein Colorimetric Assay Kit (Elabscience Biotechnology Co., Ltd., China) was used to quantify the total protein of brain cortex tissues and SY5Y cells. Samples with an equal amount of protein were loaded and separated via sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). The proteins were transferred to the appropriate polyvinylidene fluoride membrane (Millipore, USA), blocked with 5% skimmed milk for 1 h, and thens incubated with diluted antibodies at 4°C overnight. For the in vivo experiments, Drp1 (1:2000), Fis1 (1:1500), MFF (1:8000), OPA1 (1:2000), Mfn1 (1:1000), Mfn2 (1:2000), and β‐actin (1:5000) antibodies were purchased from Proteintech Group, Inc., China. Meanwhile, for the in vitro experiments, the antibodies included Drp1 (1:800; AiFang Biological, China), Fis1 (1:1500; Bioss, China), OPA1 (1:1000; AiFang Biological, China), Mfn1 (1:1000; Affinity, China), Mfn2 (1:500; AiFang Biological, China), and SENP6 (1:1000; Abcam, USA). Furthermore, MFF (1:2000), SUMO1 (1:2000), SUMO2/3 (1:500), SENP1 (1:2000), SENP2 (1:1000), SENP3 (1:800), SENP5 (1:3000), and β‐actin (1:5000) were obtained from Proteintech Group, Inc., China. Then, the membranes were washed and incubated with secondary antibodies for 2 h at room temperature. After rewashing with TBST buffer, the membranes were added ECL reagent to enable the detection of the protein bands.