Fig 1.
Expression, purification, and western blotting of Annexin B1 and B2.
(A). SDS-PAGE analysis of recombinant B1 protein expression. (B). SDS-PAGE analysis of recombinant B2 protein expression. M: Protein Marker; 1: Total protein before induction; 2: 20°C supernatant; 3: 20°C precipitation; 4: 37°C supernatant; 5: 37°C precipitation. (C). Purification of Annexin B1 by nickel agarose affinity chromatography and SDS-PAGE analysis. M: Protein marker; 1: sample; 2: outflow; 3-4: 20 mM imidazole elution component; 5: 50 mM imidazole elution component; 6: 100 mM imidazole elution component; 7: 500 mM imidazole elution component. (D). Purification of Annexin B2 by nickel agarose affinity chromatography and SDS-PAGE analysis. M: Protein marker; 1: sample; 2: outflow; 3: 20 mM imidazole elution component; 4: 50 mM imidazole elution component; 5-6: 500 mM imidazole elution component. (E). SDS-PAGE analysis of recombinant protein. M: Protein marker; 1: Annexin B1; 2: Annexin B2. (F). Recombinant protein with pig positive serum of Cysticercus cellulosae. M: Protein marker; 1: Annexin B1; 2: Annexin B2. (G). Recombinant protein with pig negative serum. (H). Recombinant protein with anti-His monoclonal antibody.
Fig 2.
Western Blotting analysis and titer determination of polyclonal antibodies.
(A). Annexin B1 with B1 polyclonal antibodies. (B). Annexin B2 with polyclonal antibodies. (C) Proteins with rabbit negative serum. M: Protein marke 1: Annexin B1; 2: Annexin B2. (D). Determination of rabbit polyclonal antibody titer.
Fig 3.
Annexin B1 and Annexin B2 localization in Cysticercus cellulosae.
Fig 4.
Annexin B1 and B2 affect the determination of APTT and PT.
The dotted area is the normal time range. Plasma added with aprotinin as positive control of APTT. Plasma lacking coagulation factor FVII as positive control of PT. The original plasma samples were used as negative controls. NS is normol saline. P<0.05 (*), P<0.01 (**), P<0.001 (***), P<0.0001 (****), ns is non-significant (n=30).
Fig 5.
Western blotting of liposome binding assay and total cellular protein.
Binding test of liposomes with different PS/PC combinations of recombinant Annexin B1 and B2. A: Annexin B1; B: Annexin B2. Numbers 1-6 represent different PS/PC binding ratio spots, which are 5/0, 4/1, 3/2, 2/3, 1/4 and 0/5 respectively.
Fig 6.
Uptake of FM1-43 dye after laser injury in media in a C2C12 cell.
(A). +Ca2+ repair group of Annexin B1. (B). +Ca2+ repair group of Annexin B2. (C). +Ca2+ repair group of ordinary cells. (D). -Ca2+ no repair group of ordinary cells. The point of the arrow is the site of the injury. (E). Results of Western blot analysis of total cell protein. M: marker; 1: Cells transfected with Annexin B1 gene; 2: Cells transfected with Annexin B2 gene; 3: Cells transfected with pCDNA3.1-His; 4: Original C2C12.
Fig 7.
Fluorescence intensity quantization of FM1-43 in cells.
(A). Group transfected with Annexin B1 gene. (B). Group transfected with Annexin B2 gene. (C). Group of original C2C12. (D). Group transfected with pCDNA3.1-His.