1. IHC localization of MASP-3 protein in joints in mice with arthritis.
IHC localization of MASP-3 protein in joints in mice with arthritis. Representative images of the joints from WT with CAIA-treated GalNAc–Luc–siRNA and GalNAc–MASP-3 siRNA. The top set of four panels from left to right (A, C, E, and G) show positive staining with biotinylated anti-mouse MASP-3 (dark brown color) (black arrow) from the liver (first panel), brain (second panel), knee joints (KJ) (third panel), and forepaws (FP) (last leftmost panel) of WT mice. The hepatocytes (H) (red arrow) present in the liver were positive for MASP-3, whereas brain (i.e., granular layer cells [GL]) were all strongly positive for MASP-3 (A and C). Many cells were positive for MASP-3 protein in the synovium (S) from KJ and FP in WT mice with CAIA treated with GalNAc–Luc–siRNA. The middle set of four panels from left to right (B, D, F, and H) show no staining (no dark brown color) used as negative controls by staining using diluent alone without primary Ab from the liver (first middle panel) and brain (second middle panel) of WT mice. There were no or few MASP-3–positive cells in the KJ (third middle panel) and FP (last middle rightmost panel) of WT mice with CAIA treated with GalNAc–MASP-3–siRNA. The single bottom panel shows no MASP-3 staining (no dark brown color) (negative control) from the liver of MASP-1/3−/− mice (I) (bottom panel) as expected. The liver, brain, KJ, and FP sections stained for MASP-3 deposition were counterstained with H&E and photographed at original magnifications ×20 or ×40 using Zeiss Observer. D1 (Axio) microscope. Joint sections were examined in duplicate from WT (n = 7) with CAIA treated with GalNAc–MASP-3–siRNA and from WT with CAIA treated with GalNAc–luciferase–siRNA, but representative pictures from each treatment groups have been shown. Red scale bar shown in (A), (B), (E), and (F), 20 μm. Red scale bar in (C), (D), (G), (H), and (I), 50 μm. CTRL, control. LUC, luciferase.
Nirmal K. Banda et al. ImmunoHorizons 2018;2:
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