Fig. 2: Protein carbonylation, ubiquitination and aggresome formation. | Communications Biology

Fig. 2: Protein carbonylation, ubiquitination and aggresome formation.

From: Immunoproteasome remodeling in senescing human macrophages reveals the loss of PA28αβ capping as a hallmark of immunosenescence

Fig. 2

a Protein carbonylation, as detected with an OxyBlot kit. Five micrograms of protein derivatized with DNPH and nonderivatized, as a negative control, were separated by SDS‒PAGE on a 12% (w/v) acrylamide gel and subjected to Western immunoblotting with an anti-DNPH antibody. Total protein was visualized as a loading control. b Ubiquitin-conjugated proteins were detected by western immunoblotting with an antibody directed against ubiquitin. Five micrograms of total protein was separated via SDS‒PAGE on a 13% (w/v) acrylamide gel. Total protein was used as a control. c Aggresomes were detected with a ProteoStat® Aggresome Detection Kit in accordance with the manufacturer’s instructions. DRAQ5 was used to visualize the nuclei (blue). Images were acquired with a confocal Leica microscope (×63 oil objective) using a standard rhodamine filter set. Scale bars 15 µm.

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