Fig. 3: Binding of AOPPs to RANK and RAGE. | Cell Death & Disease

Fig. 3: Binding of AOPPs to RANK and RAGE.

From: Age-related accumulation of advanced oxidation protein products promotes osteoclastogenesis through disruption of redox homeostasis

Fig. 3: Binding of AOPPs to RANK and RAGE.

A, B SPR binding-affinity showed AOPPs bound to RANK and RAGE. C Immunofluorescence staining showed that TRITC-AOPPs (red) co-localized with RANK (green, up) or RAGE (green, down) on the plasma membrane of BMMs. Scale bars = 5 µm. D BMMs were treated with AOPPs. Co-immunoprecipitation (IP) analysis of AOPPs and RANK or RAGE association in BMMs. Immunoblot (IB) probed using indicated antibodies. E FACS analysis of AOPPs binding to HEK293T cells transfected with RANK or RAGE. RANK increased from 56.66 ± 1.95% (red, positive cells) to 64.99 ± 2.68% (green, RANK overexpressing cells) and 73.17 ± 2.94% (blue, RAGE overexpressing cells), and the control cell (gray) was 0.86 ± 0.34%. F In BMMs, knockdown of endogenous RANK decreased TRITC-AOPPs binding from 84.50 ± 3.53% (red) to 54.72 ± 1.58% (blue, RANK knockdown cells), and 39.46 ± 2.04% (orange, RAGE knockdown cells). *p < 0.05 versus the vehicle-treated group. #p < 0.05 versus the AOPPs-treated group.

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