Anti gadd45g

The human liver cancer cells SK-HEP-1 was obtained from the Procell Life Science & Technology Co., Ltd (Wuhan, China) and was authenticated through STR genotyping. The cell lines were tested periodically for mycoplasma contamination during the experiment and were confirmed negative. Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, CA, USA) and 1% penicillin–streptomycin solution (Beyotime, Shanghai, China).
4MOD was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). In animal experiments, 0.25% polyoxyl castor oil (Yuanye, Shanghai, China) and 0.25% EtOH were used to dissolve 4MOD. A rabbit polyclonal antibody anti-GADD45G was obtained from Bioss (Beijing, China).

Seventy-six HCC and 60 paracancerous tissues were collected from the First Affiliated Hospital of Guangxi Medical University between 1 April 2019 and 1 May 2021. 4MOD-treated and 4MOD-untreated tumor tissues were obtained from the aforementioned HCC nude mouse xenografts. The primary antibody used in IHC assay was a rabbit polyclonal antibody anti-GADD45G (1:200 dilution, Bioss, Beijing, China). The secondary antibody kit (PK1006, Proteintech, Wuhan, China) was purchased from Wuhan Sanying Biotechnology. After formalin fixation and paraffin embedding, all tissues were cut into 4-μm sections, and then deparaffinized and rehydrated. Pressure cooking with Tris–EDTA buffer (PH 9.0) for 5 min was used for antigen retrieval, and then the sections were blocked with 3% H₂O₂ and 10% normal goat serum. After blocking, tissue sections were incubated with GADD45G antibody overnight at 4 °C, washed with phosphate-buffered saline (PBS), and incubated with secondary antibody at room temperature for 1 h. IHC staining score in this study were calculated using a semi-quantitative approach described in detail in previously reference [25 (link)].