Fig 1: Multiplexed immunostaining of representative control (A, D, and G), nonfibrotic (B, E, and H), and profibrotic COVID-19 hearts (C, F, I, and K), showing HSP47 (heat shock protein 47), CD163, and collagen a 1(l) (Cola1[I]) expression in myofibroblasts and macrophages.Cardiomyocytes are marked by gray (cardiac muscle troponin T [cTnT]; Abcam, number ab8295), and nuclei are visualized by royal blue 4',6-diamidino-2-phenylindole (DAPI) (fluorescent DNA stain) (Akoya Biosciences, number FP1490) staining. C, We observed increased HSP47+ (cyan blue; Santa Cruz, number SC-5293) and CD163+ (white; Leica Biosystems, number NCL-L-CD163) in COVID-19 profibrotic samples. F, Extracellular matrix deposition of Cola1(I) (red; Abcam, number ab138492) is increased in profibrotic COVID-19 samples. I, HSP47, DAPI, cTnT, and CD163 merged with Cola1(I). J, Secondary antibody (ab) alone showed no fluorescence. K, Myofibroblasts were identified by a second protein, periostin (Sigma, number HPA012306).
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