Alexa fluor 488 labeled goat anti mouse secondary antibody

Murine palatal shelf tissues of E13.5, E14.5, and E16.5 were dewaxed in xylene, and then dehydrated in ethanol. The tissues were then incubated in solutions containing specific primary antibodies: EIF3H (1:100, Santa Cruz, sc-271283), SRSF1 (1:100, Santa Cruz, sc-33652), and RBBP6 (1:100, Santa Cruz, sc-9962) (Table S3) followed by incubation in Alexa fluor 488-labeled goat anti-mouse secondary antibodies (1: 50, Beyotime, A0428).

Caco2 cells and organoids were cultured on glass-bottom dishes. After 48 h of SA11 infection, the cells and organoids were washed with PBS and fixed with 4% paraformaldehyde for 10 min. They were then blocked with a blocking buffer from Beyotime biotechnology for 60 min. Following blocking, the samples were incubated overnight at 4 °C with an anti-rotavirus antibody (ab181695) from Abcam (Cambridge, UK). Subsequently, the samples were incubated with Alexa Fluor 488-labeled goat anti-mouse secondary antibodies (Beyotime Biotechnology) at a 1:500 dilution. The nucleus was stained with DAPI (4,6-diamidino-2-phenylindole) from Beyotime biotechnology for 5 min at room temperature. Finally, the images were captured using immunofluorescence microscopy.

The washing buffer (5 $\times$ 10^− 3 м MgCl₂ and 4.5 g L^− 1 glucose in Dulbecco’s PBS) was obtained from Sigma-Aldrich (St. Louis, MO, USA), and the binding buffer was composed with yeast tRNA (0.1 mg mL^− 1, Sigma-Aldrich) and bovine serum albumin (BSA; 1 mg mL^− 1 (ThermoFisher Scientific, Waltham, MA, USA) in washing buffer, which was used to reduce background binding. Super GelRed was purchased from US Everbright® Inc (Sayreville, NJ, USA). Beijing Labor Technology Co, Ltd. provided the Cell Counting Kit-8 (CCK-8) (Beijing, China; cat. no. 21,162,196). Protein markers were purchased from Sangon Biotech Co. Ltd. (Shanghai, China). The anti-PTK7 rabbit monoclonal antibody was purchased from Abcam (Cambridge, MA, USA). The Alexa Fluor 488-labeled goat anti-mouse secondary antibody, Hoechst 33,342, lysotracker, radioimmunoprecipitation assay (RIPA) lysis buffer, phosphatase inhibitors cocktail, protease inhibitors, and annexin V-FITC Apoptosis Detection Kit (no. 011821210207) were all purchased from Beyotime Biotech Inc (Jiangsu, China). All chemicals for synthesis and purification were purchased from Energy Chemical (Shanghai, China). All tubes and plates for cell culture were purchased from NEST Biotechnology (Jiangsu, China). Unless otherwise stated, all other bioreagents were purchased from Sigma.