Mouse anti flag monoclonal antibody

Expression of the target proteins was analyzed by western blotting. The recombinant Lactobacillus were broken up by ultrasound and electrophoresis using 10% SDS-PAGE. Mouse anti-FLAG monoclonal antibody (diluted 1:1,000; Abcam, Cambridge, MA, USA) was used as the primary antibody, and horseradish peroxidase-conjugated goat anti-mouse IgG (diluted 1:5,000; ZSGB Biotech, Beijing, China) was used as the secondary antibody. Furthermore, 6aa-COE/L. reuteri and 87aa-COE/L. reuteri were cultured overnight at 37°C in MRS broth containing chloramphenicol (10 μg mL^-1). The bacteria were pelleted by centrifugation at 1500 × g for 5 min, washed three times with sterile PBS, and resuspended in sterile PBS. The suspended bacterial pellet was sequentially incubated with anti-FLAG tag mouse monoclonal antibody (diluted 1:1,000) and FITC-conjugated anti-mouse IgG secondary antibodies (diluted 1:250). Fluorescence of the target protein expressed on the surface of Lactobacillus was measured by fluorescence microscopy.

Wortmannin, dynasore, SU6655, NH₄Cl, SP600125, IPA-3, MβCD and chlorpromazine were obtained from Sigma (Sigma, MO, United States), Akti-1/2 and bafilomycin A1 were obtained from Abcam (Abcam, Cambridge, United Kingdom), nystatin and EIPA was obtained from Solarbio (Solarbio, Beijing, China).
The rabbit anti-dynamin-2, anti-clathrin heavy chain, anti-Rab5, anti-Rab7, anti-PI3K p85 (phos pho Y458) + PI3 Kinase p55 (phospho Y199), anti-Akt, anti-Src, anti-Src (phospho Y416), anti-phospho-Akt (Ser473) and anti-GAPDH monoclonal antibodies were obtained from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-JNK and anti-JNK (Thr183/Tyr185) monoclonal antibodies were obtained from Cell Signaling Technology (Cell Signaling Technology, Danvers, United States). The rabbit anti-PI3 Kinase p85 alpha polyclonal antibody, the mouse anti-Flag monoclonal antibody, goat anti-rabbit and goat anti-mouse HRP-labeled secondary antibody, Alexa fuor-488-conjugated anti-mouse, Cy™3-conjugated anti-rabbit IgG (H + L) and mounting medium with DAPI was purchased from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-caveolin-1 monoclonal antibody was obtained from Beyotime (Beyotime Biotechnology, Shanghai, China). The mouse anti-BRSV G protein monoclonal antibody was provided by China Animal Health and Epidemiology Center.