Journal: Animals : an Open Access Journal from MDPI

Article Title: Protective Effects of Recombinant Lactobacillus paracasei Expressing Porcine β-Defensin 2 Against DSS-Induced Colitis in a Murine Model

doi: 10.3390/ani16101425

Figure Lengend Snippet: The pPG-N1-pBD2/ 27-2 expressing pBD2 were identified. ( a ). Double enzyme digestion results of pPG-PPT plasmid. ( b ) PCR amplification of the N1-pBD2 fragment. ( c ) PCR identification of the pPG-N1-pBD2 plasmid. ( d ) PCR Identification results of pPG-N1-pBD2/ 27-2 . ( e ) Western blot identification of the supernatant and precipitation from pPG-N1-pBD2/ 27-2 culture. ( f ) Identification of proteins expressed in the pPG-N1-pBD2/ 27-2 . Mouse anti-flag monoclonal antibody was used as the primary antibody, and goat anti-mouse IgG labeled with FITC was used as the secondary antibody (green). Subsequently, the cell nuclei were stained with DAPI (blue), and the results were observed under an inverted fluorescence microscope. Scale bar = 50 μm.

Article Snippet: The mouse anti-flag monoclonal antibody (Abmart, Shanghai, China) was used at a dilution of 1:2000, in conjunction with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG (Invitrogen, Carlsbad, CA, USA) diluted at 1:4500, and incubated at ambient temperature for 3 h. Following this, the samples were washed three times with sterile PBS and then incubated with 4′,6′-diamino-2-phenylindole (DAPI) (Invitrogen, Carlsbad, CA, USA) for 5 min.

Techniques: Expressing, Plasmid Preparation, Amplification, Western Blot, Labeling, Staining, Fluorescence, Microscopy