anti antimouse primary  (Proteintech)

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Infectivity of lentiviruses in mesenchymal stem cells (MSCs). (a) Representative fluorescent microscopy images illustrated the expression of GFP in MSCs infected with lentivirus (scale bar = 30 μm). Quantitative analysis of the level of p38 MAPK mRNA (b) and protein expression (c) in MSCs after lentiviral transfection. **P < 0.01.

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Infection, Microscopy, Expressing, Transfection

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Downregulation of p38 MAPK does not affect the cell cycle or cellular apoptosis of mesenchymal stem cells (MSCs). (a) Representative images of the cell cycle analyzed by FACS are shown in the upper panel. (b) DNA content during different periods of the cell cycle indicated that there was no significant difference between p38-shRNA infected MSCs and NC-shRNA-infected MSCs. (c) Representative images of cell apoptosis analyzed by FACS are shown in the lower panel. Early apoptosis is shown in the right lower quadrant. (d) No significant difference was observed in early apoptosis analyses between the two different groups. P > 0.05. Cell cycle and apoptosis experiments indicated that downregulation of p38 MAPK does not affect the cell cycle or cellular apoptosis of MSCs.

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: shRNA, Infection

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Mesenchymal stem cells (MSCs) infected with p38 MAPK-shRNA promote the recovery of neurological function and a reduction in infarct volume in mice. Quantitative analysis of modified neurological severity score (mNSS) analysis (a) and foot-fault tests (b) in mice of two groups on day 14 (*P < 0.05). (c) Representative T2-weighted images of MSC-transplanted mice in two groups on day 21 are shown in the lower panel. Infarct regions are outlined by yellow dotted line. (d) Quantitative analysis of delta infarct volume, that is, the difference in the infarction volume at day 0 and day 21 showed a significant reduction in infarct volume in mice treated with p38 MAPK-shRNA infected MSCs (*P < 0.05).

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Infection, shRNA, Modification

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Mesenchymal stem cells (MSCs) infected with p38 MAPK-shRNA increase axonal remodeling in mice. (a) Diffusion weighted imaging reconstruction showed that the corticospinal tract on the bilateral side. Region of interest was set at the bilateral internal capsule. Quantitative analysis of fractional anisotropy (FA) value (b) and the number (c) and length (d) of fiber tracts in the ipsilateral internal capsule showed significant differences between the two groups. (e) Representative myelin basic protein (MBP) immunohistochemical staining of the ipsilateral internal capsule region are shown (scale bar = 100 μm). (f) Statistical analysis of MBP staining in two groups showed significantly increased number of MBP-positive cells in the ipsilateral internal capsule in the p38-shRNA treated MSCs group. *P < 0.05.

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Infection, shRNA, Diffusion-based Assay, Imaging, Immunohistochemical staining, Staining

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Downregulation of p38 MAPK decreases cellular senescence in mesenchymal stem cells (MSCs). (a) Senescence-associated β-galactosidase staining assays of MSCs in two different groups are shown. Senescent cells exhibit high expression of SA-β-gal in blue staining (scale bar = 30 μm). (b) Quantitative analysis of the number of senescent MSCs showed significant differences between the two groups (**P < 0.01), indicating that downregulation of p38 MAPK decreases cellular senescence in MSCs.

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Staining, Expressing

Journal: Experimental Biology and Medicine

Article Title: Mesenchymal stem cells with p38 mitogen-activated protein kinase interference ameliorate mouse ischemic stroke

doi: 10.1177/15353702231220663

Figure Lengend Snippet: Mesenchymal stem cells (MSCs) infected with p38 MAPK-shRNA promote neurogenesis and decrease apoptosis in mice. (a) Schematic representation of ischemic brain area and sampling areas. TUNEL assay samples are from the area in the black frame, while DCX/Brdu staining samples are from the area in the red frame. (b and c) TUNEL assay indicates decreased cell apoptosis around the ischemia border region in the p38-shRNA group (scale bar = 30 μm). (d and e) Immunofluorescence staining showed more DCX/Brdu double-positive cells in the iplsilateral subventricular zone in the p38-shRNA group than in the control group (scale bar = 30 μm). *P < 0.05; **P < 0.01.

Article Snippet: The proteins were separated in a 12% separation gel and then transferred (wet transfer at 300 mA for 70 min) to a membrane, which was blocked in 5% non-fat milk for 1 h. The membranes were then incubated with rabbit antimouse p38 MAPK primary antibody (1:1000, Cell signaling Technology, Danvers, MA, USA) and GAPDH antibody (1:2000, Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Infection, shRNA, Sampling, TUNEL Assay, BrdU Staining, Immunofluorescence, Staining, Control