Fig. 6

Specificity of the 1A8, 4F5, and R&D972207 antibodies for DEFA5. A Cell lysates from HEK293T cells expressing recombinant DEFA1, DEFA5, and DEFA6 α defensin family members were analyzed by immunoprecipitation and Western blotting. A1–A3 Subfigures represent the detection of recombinant DEFA1, DEFA5, and DEFA6 in HEK293T cell lysates using 1A8, 4F5, and R&D972207 antibodies, respectively. Lanes 1, 3, and 5: Negative control IP with mouse IgG for recombinant DEFA1, DEFA5, and DEFA6 HEK293T cell lysates, respectively. Lanes 2, 4, and 6: IP with sc-53997 antibody for DEFA1, DEFA5, and DEFA6 HEK293T cell lysates, respectively. Immunoprecipitants were used in Western blot and each blot was incubated with 1A8 antibody (A1), 4F5 antibody (A2), and R&D972207 antibody (A3). B Densitometric analysis of the precipitated DEFA5 protein. Bars represent the mean intensity of the DEFA5 band from two independent experiments relative to the corresponding IgG control. C (C1–C3) Subfigures represent the detection of control lysates from HEK293T cells transfected with empty expression vectors for DEFA1, DEFA5, and DEFA6. Lanes 1, 3, and 5: Negative control IP with mouse IgG for lysates from HEK293T cells transfected with empty DEFA1, DEFA5, and DEFA6 expression vectors, respectively. Lanes 2, 4, and 6: IP with sc-53997 antibody for lysates from HEK293T cells transfected with empty DEFA1, DEFA5, and DEFA6 expression vectors, respectively. Immunoprecipitates were used in Western blot and each blot was incubated with 1A8 (C1), 4F5 (C2), and R&D972207 (C3) antibodies. D Cell lysates from HEK293T cells expressing recombinant DEFA1, DEFA5, and DEFA6 α defensin family members were analyzed by standard Western blotting. Detection of GAPDH was used as the loading control