Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 1. B Representative bands and ( C – G ). quantitative analyses of endogenous MFGE8 integrin β3, p-Akt, Akt, cyclin D1 and HPCA in left hippocampus after SAH. The MGFE8, integrin β3 and p-Akt are increased and peaked at 24 h after SAH, while cyclin D1 and HPCA show the opposite trend. ** P < 0.01 vs sham group. Error bars were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. H Double immunofluorescence staining of MFGE8 (green) with DCX (immature neuron marker, red), Iba-1(microglia marker, red) and GFAP (astrocytes marker, red) in hippocampus at 24 hr after SAH. Red box indicated the hippocampus in the brain slice. n = 2 per group. Scale bar, 50 μm. DCX doublecortin, Iba-1 ionized calcium binding adapter molecule-1, GFAP glial fibrillary acidic protein, DAPI diamidino phenylindole.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Double Immunofluorescence Staining, Marker, Slice Preparation, Binding Assay

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 2. B Double immunofluorescence staining with DCX (red) and BrdU (green) in hippocampus between sham, SAH + vehicle and SAH + rhMFGE8 group as well as SAH + Scr siRNA, SAH + MFGE8 siRNA group. Arrows indicated the DCX positive and BrdU positive cells. C Quantitative analyses of DCX positive BrdU positive cells in three groups. The DCX positive BrdU positive cells are decreased in SAH+vehicle group compared to sham, which are reversed by rhMFGE8 treatment. The cells are also decreased in SAH + MFGE8 siRNA group compared to SAH + Scr siRNA group. The Red box indicated the hippocampus in the brain slice. ** P < 0.01 vs sham group, ## P < 0.01 vs SAH + vehicle group, $$ P < 0.01 vs SAH + Scr siRNA group. Error bars were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 3 per group. Scale bar, 100 μm. BrdU bromodeoxyuridine.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Double Immunofluorescence Staining, Slice Preparation

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 4. B Modified Garcia score and beam balance score show that rhMFGE8 treatment improves the scores, with the effect reversed by int-β3 siRNA and PI3K/Akt inhibitor. C Representative western blot bands and ( D – K ). Quantitative analyses of MFGE8, int β3, PI3K, p -Akt, Akt, mTOR, cyclin D1, HPCA and DCX at 24 hr after SAH. ** P < 0.01 vs sham group; ## P < 0.01 vs . SAH+vehicle group; $$ P < 0.01 vs. SAH + rhMFGE8+ scr siRNA; && P < 0.01 vs. SAH+rhMFGE8 + vehicle . Data were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Modification, Western Blot

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 1. B Representative bands and ( C – G ). quantitative analyses of endogenous MFGE8 integrin β3, p-Akt, Akt, cyclin D1 and HPCA in left hippocampus after SAH. The MGFE8, integrin β3 and p-Akt are increased and peaked at 24 h after SAH, while cyclin D1 and HPCA show the opposite trend. ** P < 0.01 vs sham group. Error bars were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group. H Double immunofluorescence staining of MFGE8 (green) with DCX (immature neuron marker, red), Iba-1(microglia marker, red) and GFAP (astrocytes marker, red) in hippocampus at 24 hr after SAH. Red box indicated the hippocampus in the brain slice. n = 2 per group. Scale bar, 50 μm. DCX doublecortin, Iba-1 ionized calcium binding adapter molecule-1, GFAP glial fibrillary acidic protein, DAPI diamidino phenylindole.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Double Immunofluorescence Staining, Marker, Slice Preparation, Binding Assay

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 2. B Double immunofluorescence staining with DCX (red) and BrdU (green) in hippocampus between sham, SAH + vehicle and SAH + rhMFGE8 group as well as SAH + Scr siRNA, SAH + MFGE8 siRNA group. Arrows indicated the DCX positive and BrdU positive cells. C Quantitative analyses of DCX positive BrdU positive cells in three groups. The DCX positive BrdU positive cells are decreased in SAH+vehicle group compared to sham, which are reversed by rhMFGE8 treatment. The cells are also decreased in SAH + MFGE8 siRNA group compared to SAH + Scr siRNA group. The Red box indicated the hippocampus in the brain slice. ** P < 0.01 vs sham group, ## P < 0.01 vs SAH + vehicle group, $$ P < 0.01 vs SAH + Scr siRNA group. Error bars were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 3 per group. Scale bar, 100 μm. BrdU bromodeoxyuridine.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Double Immunofluorescence Staining, Slice Preparation

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Experimental design, animal groups and time frame of the analyses in experiment 4. B Modified Garcia score and beam balance score show that rhMFGE8 treatment improves the scores, with the effect reversed by int-β3 siRNA and PI3K/Akt inhibitor. C Representative western blot bands and ( D – K ). Quantitative analyses of MFGE8, int β3, PI3K, p -Akt, Akt, mTOR, cyclin D1, HPCA and DCX at 24 hr after SAH. ** P < 0.01 vs sham group; ## P < 0.01 vs . SAH+vehicle group; $$ P < 0.01 vs. SAH + rhMFGE8+ scr siRNA; && P < 0.01 vs. SAH+rhMFGE8 + vehicle . Data were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 6 per group.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Modification, Western Blot

Journal: Cell Death Discovery

Article Title: MFGE8 promotes adult hippocampal neurogenesis in rats following experimental subarachnoid hemorrhage via modifying the integrin β3/Akt signaling pathway

doi: 10.1038/s41420-024-02132-x

Figure Lengend Snippet: A Immunohistochemical staining of DCX and ( B ). quantitative analyses of DCX relative density and DCX positive cell in the hippocampus. Red box indicated the hippocampus in the brain slice. ** P < 0.01 vs sham group; ## P < 0.01 vs . SAH + vehicle group; $$ P < 0.01 vs. SAH + rhMFGE8+ scr siRNA; && P < 0.01 vs. SAH + rhMFGE8 + vehicle . Data were represented as mean ± SD. One-way ANOVA, Tukey’s test, n = 3 per group.

Article Snippet: MFGE8 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA), integrin β3 siRNA (10 μl, concentration of 5000 IFU/μl, SantaCruz, USA) or scrambled siRNA (10 μl, concentration 5000 IFU/μl, Santa Cruz, USA) were administered by i.c.v at 48 hr before SAH [ ].

Techniques: Immunohistochemical staining, Staining, Slice Preparation