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. 2014 Apr 7;5(9):2750–2760. doi: 10.18632/oncotarget.1887

Figure 4. 4E1 recognizes a conformational epitope localized on a region overlapping D1 and DX domains of CD93.

Figure 4

A: Human Lenti-X 293T cells were transiently transfected with empty vector (mock) or a construct expressing human CD93 full-length. Cell extracts were immunoprecipitated with 4E1 and immunoprecipitates were analyzed by Western blotting with anti-CD93 antibodies (H190). Equal loading of cell lysates was confirmed by using anti-β-actin antibodies. B: The schematic diagram illustrates the deletion mutants of the CD93 extracellular domains fused with a 6X Myc tag. The deletion mutants were designated in accordance with Wu and colleagues [15]. LS, signal peptide; CTLD, C-type lectin-like domain; X, domain of unknown structural function; EGF-like, EGF-like repeats; Mucin mucin-like domain; Myc, 6X Myc tag. C: Lenti-X 293T cells were transiently transfected to generate recombinant CD93 domain proteins fused with a 6X Myc tag. For each different deletion mutant two different cDNA constructs were transfected (#1 and #2). Cell extracts were analyzed by immunoblotting using anti-Myc antibodies. Anti-β-actin antibodies were used to confirm equal loading. D: Cell lysates from cells transfected as in C, were subjected to immunoprecipitation analyses by using 4E1. Immunoprecipitates were analyzed by Western blotting with anti-Myc antibodies.