Figure 9.
AXT107 cross-links with integrins α5 and β3
P16 mice reared in room air (1 set of 7 mice) or OIR (2 sets of 7 mice) were injected intravitreally with sulfo-SBED-labeled AXT107, and their retinas were isolated after 24h. Bound peptide was then cross-linked using UV reactive chemistry, and the tissue was lysed. Each lysate was then separated into three types of samples called input, pull-down, or flowthrough for the lysates before pull-down, components that associated with the beads, and components that did not associate with the beads, respectively.
(A–D) Input samples from each treatment set were separated by SDS-PAGE and analyzed by western blot for the levels of integrins α5 (A), αv (B), β1 (C), and β3 (D) with β-tubulin as a loading control.
(E–G) Input, pull-down, and flowthrough samples from each of the three datasets resolved by SDS-PAGE and analyzed for integrins α5 (E), αv (F), and β1 and β3 (G) with β-tubulin as a loading control. Input samples for integrin α5 are an extended cropping of the gel used for panel A. Conversely, the input samples in panels F and G are included to allow same gel comparisons of the three different samples but were rerun for panels B, C, and D to address an overloading artifact in lane 1. Owing to differences in molecular weights and staining intensity, integrins β1 and β3 were stained serially on the same blot.