Figure 5.

Figure 5. Ultrastructural damage, calcifications and macrophages present in skeletal muscle from cachectic PDAC patients. A–C) Representative electron micrographs of skeletal muscle from non-cancer control patients (n = 2) and cachectic PDAC patients (n = 2) following transmission electron microscopy. Areas of ECM deposition are indicated by black arrows. D–K) Representative skeletal muscle sections from non-cancer control patients (D, H) and cachectic PDAC patients (E–G, I–K) stained with Alizarin Red S to label calcium deposition (stains red). Calcium deposits localized inside (*) and at the periphery (white arrows) of muscle fibers, in the extracellular matrix (white arrowheads) and in blood vessels walls (v) are noted. Scale bar: 50 µm. L) The percent of total muscle area positive for calcium deposition in non-cancer controls (n = 3) versus cachectic PDAC patients (n = 6), expressed as the mean ± the SEM (P = .0238, Mann-Whitney U test). M–O) Representative skeletal muscle sections from a non-cancer control patient (M) and cachectic PDAC patients (M, O) immunostained with a CD68 antibody to label macrophages (brown staining, black arrows). Scale bar: 200 µm. P) Staining of a serial muscle section with H&E to demonstrate the localization of CD68+ macrophages in cachectic muscle. Areas of collagen (light pink staining, white arrows), lipid (white arrowheads) and muscle fibers (black astericks) are indicated. Q) The average number of CD68+ macrophages in muscle of cachectic PDAC patients compared to non-cancer control subjects (P = .0303, Mann-Whitney U test). Data represent mean ± SEM, from n = 5 non-cancer controls and n = 7 cachectic PDAC patients. CON = controls; PDAC = pancreatic ductal adenocarcinoma.