integrin β3 sc-29375  (Santa Cruz Biotechnology)

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

Journal: Nature Communications

Article Title: Control cell migration by engineering integrin ligand assembly

doi: 10.1038/s41467-022-32686-2

Figure Lengend Snippet: a Schematic illustration of precise control of integrin ligand presentation on nanofilaments via peptide assembly. b TEM images of nanofilaments obtained via molecular self-assembly and co-assembly of FFFIKLLI (100 μM) and FFF at various ratios, and the estimated molecular packing structures. IKLLI motif is presented in blue and FFF motif is presented in pink. The scale bars represent 200 nm. Three independent experiments were performed. c Zoom-in SEM images (false color) of HuH-7 cell edge and apical membrane after 3-day incubations. FFFIKLLI was maintained at a concentration of 100 μM. The cell body is highlighted in pink, while the nanofilaments are highlighted in blue. The scale bars represent 300 nm.

Article Snippet: Integrin β1 shRNA plasmid (#sc-29375-SH), Integrin α3 shRNA plasmid (#sc-35684-SH), Integrin α6 shRNA plasmid (#sc-43129-SH), and control shRNA plasmid-A (#sc-108060) were purchased from Santa Cruz Biotechnology for knockdown of the target integrins.

Techniques: Control, Membrane, Concentration Assay

Journal: Nature Communications

Article Title: Control cell migration by engineering integrin ligand assembly

doi: 10.1038/s41467-022-32686-2

Figure Lengend Snippet: a Time-lapse series showing actin cytoskeleton (grey) and paxillin (green) in HuH-7 cells expressing mRuby-Lifeact-7 and mGFP-paxillin upon the treatment of FFFIKLLI (100 μM) for 12 hr. Scale bars represent 2 μm. Three independent experiments were performed. b F-actin phalloidin staining (magenta), integrin β1 (cyan) and paxillin (yellow) immunofluorescence in HuH-7 cell after 12 h treatment of FFFIKLLI (100 μM). Scale bar represents 2 μm. c Fluorescence intensity distribution profile of integrin β1, paxillin, and actin cytoskeleton along the yellow line on merged image of b . d 12 h time course Rac1 activity of HuH-7 cells with or without the treatment of FFFIKLLI (100 μM). Rac1 activity was measured by FRET. n = 6 cells (Ctrl) or 5 cells (FFFIKLLI). Symbols represent the mean FRET/CFP emission ratio ± s.d. e Representative FRET/CFP ratio images of HuH-7 cells expressing RaichuEV-Rac1 with or without the treatment of FFFIKLLI (100 μM) at the indicated time points and coded according to a pseudo-color scale, which ranges from yellow to purple with an increase in Rac1 activity. Scale bars represent 20 μm. Source numerical data are available in source data.

Article Snippet: Integrin β1 shRNA plasmid (#sc-29375-SH), Integrin α3 shRNA plasmid (#sc-35684-SH), Integrin α6 shRNA plasmid (#sc-43129-SH), and control shRNA plasmid-A (#sc-108060) were purchased from Santa Cruz Biotechnology for knockdown of the target integrins.

Techniques: Expressing, Staining, Immunofluorescence, Fluorescence, Activity Assay