Fig 1: Co-localization of iNOS, Iba-1, and GFAP in MCAO mouse brain section. The mice were sacrificed 24 h after MCAO. Immunofluorescence stain was used to test the co-localization of iNOS, Iba-1, and GFAP in the MCAO mouse brain section. The iNOS (green) was mainly expressed in the infarct area and not expressed in the healthy area in the contralateral hemisphere. The iNOS protein was co-localized with GFAP (blue), but not with Iba-1 (red). Antibody: mouse anti-iNOS (1:200, Cat#ab49999, Abcam), anti-mouse Alex 488 conjugated secondary antibody (1:200, Cat#ab150117, Abcam), goat anti-GFAP (1:200, Cat#ab53554, Abcam), anti-goat Alex 647 conjugated secondary antibody (1:200, Cat#ab150135, Abcam), rabbit anti-Iba-1 (1:400, Cat#016-20001, Wako), anti-rabbit Alex 594 conjugated secondary antibody (1:200, Cat#ab150064, Abcam)
Fig 2: Inhibition of OGD induced iNOS expression in LCN2 knockout astrocytes. The iNOS and GFAP were co-localized in primary cultured wild-type astrocytes after oxygen-glucose deprivation. iNOS did not express in LCN2 knockout astrocytes after oxygen-glucose deprivation, scale bar 200 µm. Antibody: rabbit anti-iNOS (1:200, Cat#ab17945, Abcam), anti-rabbit Alex 488 conjugated secondary antibody (1:200, Cat#ab150073, Abcam), goat anti-GFAP (1:200, Cat#ab53554, Abcam), anti-goat Alex 594 conjugated secondary antibody (1:200, Cat#ab150132, Abcam)
Fig 3: Increase of MCAO induced iNOS expression in the brain section of LCN2-lentivirus-transfected mice. The iNOS and GFAP were co-localized by immunofluorescence on brain slices of LCN2-lentivirus transfected MCAO mice. a LCN2 transfected mice had higher iNOS expression in infarcted cortex area than negative controls, scale bar 20 µm. b LCN2 transfected mice also had higher iNOS expression in the infarcted hippocampus than controls, scale bar 20 µm. Antibody: rabbit anti-iNOS (1:200, Cat#ab17945, Abcam), anti-rabbit Alex 488 conjugated secondary antibody (1:200, Cat#ab150073, Abcam), goat anti-GFAP (1:200, Cat#ab53554, Abcam), anti-goat Alex 594 conjugated secondary antibody (1:200, Cat#ab150132, Abcam)
Fig 4: Increase of OGD induced iNOS expression in LCN2-lentivirus transfected astrocytes. Astrocytes were transfected by LVLCN2 for 6 h and treated by OGD 24 h later. Astrocytes were fixed at 24 h after 6 h OGD. The immunofluorescence showed that iNOS expression was increased in LCN2 overexpressed astrocytes after oxygen-glucose deprivation, scale bar 20 µm. Antibody: rabbit anti-iNOS (1:200, Cat#ab17945, Abcam), anti-rabbit Alex 488 conjugated secondary antibody (1:200, Cat#ab150073, Abcam), goat anti-GFAP (1:200, Cat#ab53554, Abcam), anti-goat Alex 594 conjugated secondary antibody (1:200, Cat#ab150132, Abcam)
Fig 5: Inhibition of MCAO induced iNOS expression in brain section of LCN2-knockout mice. The iNOS and GFAP were co-localized by immunofluorescence on brain slices of LCN2-knockout MCAO mice. a LCN2-knockout mice did not express iNOS in the infarcted cortex, scale bar 20 µm. b LCN2-knockout mice had no iNOS expression in the infarcted hippocampus, scale bar 20 µm. Antibody: rabbit anti-iNOS (1:200, Cat#ab17945, Abcam), anti-rabbit Alex 488 conjugated secondary antibody (1:200, Cat#ab150073, Abcam), goat anti-GFAP (1:200, Cat#ab53554, Abcam), anti-goat Alex 594 conjugated secondary antibody (1:200, Cat#ab150132, Abcam)
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