Immunohistochemistry kit

IHC staining was performed using an immunohistochemistry kit (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) following the manufacturer’s instructions. A rabbit polyclonal primary antibody against C1GALT1 (HPA011294; Atlas Antibodies) was employed. After titrated the dilution, we used an antibody dilution of 1:100 for IHC. Images were captured using Aperio Digital Pathology Slide Scanners (Leica Microsystems, Inc.). The IHC staining evaluation of C1GALT1 was conducted independently by two pathologists, and the stain intensity was scored as follows: 0 (negative); 1 (weak); 2 (moderate); and 3 (strong). High expression of C1GALT1 was defined as a score of 3 in IHC staining, while low expression of C1GALT1 was defined as a score of <3 in IHC staining.

One-inch disposable sterile acupuncture needles (diameter: 0.3 mm × 25 mm) (Suzhou Medical Instrument Factory, Suzhou, China), KWD-808I versatile pulse electronic acupuncture electrotherapy machine (Suzhou Universal Acupuncture Medical Devices Co., Ltd., Suzhou, China), and CJ-2F medical clean bench (Suzhou City FengShi Animals Equipment Co., Suzhou, China) were used for the study. For the study, DSC-RX100M2 digital camera (Sony Company, Japan), Bx-70 microscopy imaging system (Olympus Company, Suzhou, China), XS-200 binocular microscopy (Jiangnan Optics Co., Ltd., Nanjing, China), chloral hydrate (Aladdin Reagent Shanghai Co., Ltd., Shanghai, China), dimethyl sulfoxide (DMSO) (Sigma, Co., St. Louis, MO, USA), ERK inhibitor PD98059 (Sigma, Co., St. Louis, MO, USA), immunohistochemistry kit (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China), and TUNEL test kit (KGI Nanjing Biological Technology Development Co., Ltd., Nanjing, China) were used.

Bleomycin A2: Nippon Kayaku Co., Ltd., batch number: Y60520.
Primary antibodies were mouse anti-human p65, IκB-α and α-SMA monoclonal IgG antibodies: bought from Santa Cruz Biotechnology, Inc. The secondary antibody was biotin-labeled goat anti-mouse IgG. Immunohistochemistry kit was purchased from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. P65 and IκB-α in situ hybridization kits: purchased from Wuhan Boster Biological Technology Co., Ltd.