Extraction:Article Title: Exacerbated VEGF up-regulation accompanies diabetes-aggravated hemorrhage in mice after experimental cerebral ischemia and delayed reperfusion.
Article Snippet: Immunoreactivities were detected using enhanced chemiluminescence reagents (RPN2106, GE Healthcare, Buckinghamshire, UK or K-12042-D10, Advansta, CA, USA) and light-sensitive films (47410 19291, Fujifilm, Tokyo, Japan), quantified using ImageJ (U.S. National Institutes of Health, Bethesda, MD, USA), and normalized using an endogenous protein (actin or α-tubulin), except for phospho (p)-Akt, p-Erk1/2, and p-p38 MAPK, which were normalized with their respective total expressions. .. After mixing lysates of infarct and penumbra areas of brain slices two to four with ice-cold RNAiso plus (9109, Takara Bio Inc., Japan) at a ratio of 1:4, total RNA was extracted by the phenol:chloroform extraction method, and cDNA was prepared from 2 μg of the extracted RNA (SuperScript VILO; 11754050, Life Technologies, USA), both according to the manufacturers’ instructions. .. Real-time PCR reactions were performed using the StepOnePlus system and SYBR Green technology (4385610, Life Technologies) with the primers listed in Additional Table 2.
Immunolabeling:Article Title: Transplantation of human neural stem cells repairs neural circuits and restores neurological function in the stroke-injured brain
Article Snippet: Their brains were fixed in 2% paraformaldehyde with 3% saccharose for 30 minutes, dehydrated with 30% saccharose overnight, embedded in cryomatrix, and cut into 15-μm coronal slices using a microtome (CM-1950, Leica, Heidelberg, Germany). .. To identify whether the transplanted hNSCs survived after 12 weeks and to determine their differentiation status within the brain, the brain slices (every eighth sections) were immunolabeled using the following primary antibodies: Stem121 (mouse monoclonal antibody, 1:500, Takara, Kyoto, Japan, Cat# Y40410, RRID: AB_2801314) for human cell labeling to assess grafted human cell bodies and double-labeling for neural cell markers, including Tuj-1 (rabbit polyclonal antibody, 1:200, Abcam, Cat# ab18207, RRID: AB_444319) for immature and mature neurons; microtubule associated protein 2 (MAP2; chicken polyclonal antibody, 1:200, Abcam, Cat# ab5392, RRID: AB_2138153) for mature neurons; NG2 (rabbit polyclonal antibody, 1:300, Abcam, Cat# ab83178, RRID: AB_10672215) to label oligodendrocytes, and GFAP (rabbit monoclonal antibody, 1:200, Abcam, Cat# ab33922, RRID: AB_732571) to label astrocytes. .. Choline acetyltransferase (ChAT; rabbit monoclonal antibody, 1:200, Abcam, Cat# ab181023, RRID: AB_2687983) was used for staining mature excitatory motor neurons, and gamma-aminobutyric acid (GABA; rabbit polyclonal antibody, 1:200, Sigma-Aldrich, Cat# A2052, RRID: AB_477652) for staining GABAergic inhibitory neurons.
Article Title: Transplantation of human neural stem cells repairs neural circuits and restores neurological function in the stroke-injured brain
Article Snippet: Their brains were fixed in 2% paraformaldehyde with 3% saccharose for 30 minutes, dehydrated with 30% saccharose overnight, embedded in cryomatrix, and cut into 15-μm coronal slices using a microtome (CM-1950, Leica, Heidelberg, Germany). .. To identify whether the transplanted hNSCs survived after 12 weeks and to determine their differentiation status within the brain, the brain slices (every eighth sections) were immunolabeled using the following primary antibodies: Stem121 (mouse monoclonal antibody, 1:500, Takara, Kyoto, Japan, Cat# Y40410, RRID: AB_2801314) for human cell labeling to assess grafted human cell bodies and double-labeling for neural cell markers, including Tuj-1 (rabbit polyclonal antibody, 1:200, Abcam, Cat# ab18207, RRID: AB_444319) for immature and mature neurons; microtubule associated protein 2 (MAP2; chicken polyclonal antibody, 1:200, Abcam, Cat# ab5392, RRID: AB_2138153) for mature neurons; NG2 (rabbit polyclonal antibody, 1:300, Abcam, Cat# ab83178, RRID: AB_10672215) to label oligodendrocytes, and GFAP (rabbit monoclonal antibody, 1:200, Abcam, Cat# ab33922, RRID: AB_732571) to label astrocytes. .. Choline acetyltransferase (ChAT; rabbit monoclonal antibody, 1:200, Abcam, Cat# ab181023, RRID: AB_2687983) was used for staining mature excitatory motor neurons, and gamma-aminobutyric acid (GABA; rabbit polyclonal antibody, 1:200, Sigma-Aldrich, Cat# A2052, RRID: AB_477652) for staining GABAergic inhibitory neurons.
Labeling:Article Title: Transplantation of human neural stem cells repairs neural circuits and restores neurological function in the stroke-injured brain
Article Snippet: Their brains were fixed in 2% paraformaldehyde with 3% saccharose for 30 minutes, dehydrated with 30% saccharose overnight, embedded in cryomatrix, and cut into 15-μm coronal slices using a microtome (CM-1950, Leica, Heidelberg, Germany). .. To identify whether the transplanted hNSCs survived after 12 weeks and to determine their differentiation status within the brain, the brain slices (every eighth sections) were immunolabeled using the following primary antibodies: Stem121 (mouse monoclonal antibody, 1:500, Takara, Kyoto, Japan, Cat# Y40410, RRID: AB_2801314) for human cell labeling to assess grafted human cell bodies and double-labeling for neural cell markers, including Tuj-1 (rabbit polyclonal antibody, 1:200, Abcam, Cat# ab18207, RRID: AB_444319) for immature and mature neurons; microtubule associated protein 2 (MAP2; chicken polyclonal antibody, 1:200, Abcam, Cat# ab5392, RRID: AB_2138153) for mature neurons; NG2 (rabbit polyclonal antibody, 1:300, Abcam, Cat# ab83178, RRID: AB_10672215) to label oligodendrocytes, and GFAP (rabbit monoclonal antibody, 1:200, Abcam, Cat# ab33922, RRID: AB_732571) to label astrocytes. .. Choline acetyltransferase (ChAT; rabbit monoclonal antibody, 1:200, Abcam, Cat# ab181023, RRID: AB_2687983) was used for staining mature excitatory motor neurons, and gamma-aminobutyric acid (GABA; rabbit polyclonal antibody, 1:200, Sigma-Aldrich, Cat# A2052, RRID: AB_477652) for staining GABAergic inhibitory neurons.
Article Title: Transplantation of human neural stem cells repairs neural circuits and restores neurological function in the stroke-injured brain
Article Snippet: Their brains were fixed in 2% paraformaldehyde with 3% saccharose for 30 minutes, dehydrated with 30% saccharose overnight, embedded in cryomatrix, and cut into 15-μm coronal slices using a microtome (CM-1950, Leica, Heidelberg, Germany). .. To identify whether the transplanted hNSCs survived after 12 weeks and to determine their differentiation status within the brain, the brain slices (every eighth sections) were immunolabeled using the following primary antibodies: Stem121 (mouse monoclonal antibody, 1:500, Takara, Kyoto, Japan, Cat# Y40410, RRID: AB_2801314) for human cell labeling to assess grafted human cell bodies and double-labeling for neural cell markers, including Tuj-1 (rabbit polyclonal antibody, 1:200, Abcam, Cat# ab18207, RRID: AB_444319) for immature and mature neurons; microtubule associated protein 2 (MAP2; chicken polyclonal antibody, 1:200, Abcam, Cat# ab5392, RRID: AB_2138153) for mature neurons; NG2 (rabbit polyclonal antibody, 1:300, Abcam, Cat# ab83178, RRID: AB_10672215) to label oligodendrocytes, and GFAP (rabbit monoclonal antibody, 1:200, Abcam, Cat# ab33922, RRID: AB_732571) to label astrocytes. .. Choline acetyltransferase (ChAT; rabbit monoclonal antibody, 1:200, Abcam, Cat# ab181023, RRID: AB_2687983) was used for staining mature excitatory motor neurons, and gamma-aminobutyric acid (GABA; rabbit polyclonal antibody, 1:200, Sigma-Aldrich, Cat# A2052, RRID: AB_477652) for staining GABAergic inhibitory neurons.
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