Affinipure goat anti mouse igg

Colon sections were incubated with anti‐LC3II (1:100, #2775, CST) and anti‐GRP78 (1:100, 66574‐1‐lg, Protein Tech) antibodies. Secondary antibodies (FITC‐conjugated AffiniPure goat anti‐mouse IgG, BA1101, TRITC‐conjugated AffiniPure goat anti‐rabbit IgG, BA1090, Boster, USA) were used in this study.
Cells were incubated with anti‐LC3II (1:500). Nuclei were stained with DAPI (1155MG010, Biofroxx, Germany). Images were obtained using a LEICA SP8 confocal microscope (LEICA, Germany). And the fluorescence intensity was quantified using ImageJ software (Fiji, ImageJ).

The CHMm and CHMp were inoculated overnight in six-well cell culture plates and incubated for 24 h, 48 h, and 72 h with the addition of medium containing varying concentrations of matrine, and a control group (with the addition of cell maintenance medium) was set up. After the culture was finished, cells were collected in EP tubes using cell scrapers, and total cellular proteins were extracted using the same method as in 2.5, and protein concentrations were determined using a BCA kit, followed by SDS-PAGE separation of the protein samples, transferring the proteins to poly (vinylidene fluoride) PVDF membranes, closing them with 5% skim milk for 2 h, adding a 1:500 dilution of mouse antibody-BTF3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) solution for overnight treatment, GAPDH (Proteintech, Wuhan, China) was used as an internal reference, and then adding HRP conjugated affiniPure goat anti-mouse IgG (Boster, Wuhan, China) for 1 h. After darkroom exposure and film scanning, image J 1.42q software was used for analysis and processing.