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Fig. 1 | Acta Neuropathologica Communications

Fig. 1

From: New insights into the protein aggregation pathology in myotilinopathy by combined proteomic and immunolocalization analyses

Fig. 1

Workflow of our combined laser microdissection and label-free proteomic approach. Frozen skeletal muscle sections from myotilinopathy patients were immunostained using a primary antibody directed against myotilin in order to detect protein aggregation in affected muscle fibers. Protein aggregates (“aggregate samples”) and aggregate-free areas in fibers with a normal appearance (“control samples”) were collected from the same section by laser microdissection (LMD). After tryptic in-solution digestion of extracted proteins, protein identification was performed by LC-ESI-MS/MS-analysis. Proteins with an over-representation in aggregate samples compared to control samples were identified by label-free relative quantification (spectral index calculation). For validation of proteomic data, serial cryosections of muscle samples from myotilinopathy patients were immunostained with primary antibodies directed against selected proteins identified as over-represented in aggregate samples

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