Journal: Molecular Oncology

Article Title: Downregulation of miR‐326 and its host gene β‐arrestin1 induces pro‐survival activity of E2F1 and promotes medulloblastoma growth

doi: 10.1002/1878-0261.12800

Figure Lengend Snippet: Biological effects of ectopic expression of miR‐326 and ARRB1 in MB CSCs. (A–C) MB CSCs were assayed 48 h after transfection with miR‐326 and ARRB1 , individually or combined. Mock‐transfected cells served as controls. Ectopic expression of ARRB1 and/or miR‐326 in these cells (A) reduced proliferation (MTT assay) (24 h: vs. CTRL: miR‐326+ P = 0.0072; ARRB1‐HA+ P = 0.0245; miR‐326+ARRB1‐HA+ P = 0.0015; 48 h: vs. CTRL: miR‐326+ P = 0.001; ARRB1‐HA+ P = 0.0005; miR‐326+ARRB1‐HA+ P < 0.0001), (B) diminished the frequency of oncosphere‐forming cells (vs. miR‐326‐ARRB1‐HA−: miR‐326+ P = 0.001; ARRB1‐HA+ P = 0.0005; miR‐326+ARRB1‐HA+ P = 0.0003), (C) decreased NANOG expression and increased the expression of PARP‐C (miR‐326 levels: miR‐326+ P < 0.0001; miR‐326+ARRB1‐HA+ P = 0.0002 vs. miR‐326‐ARRB1‐HA−). (D) qRT‐PCR revealed significantly increased expression of neuronal and glial differentiation markers ( βIII tubulin and GFAP , respectively) only in MB CSCs overexpressing miR‐326 (alone or with ARRB1 ) (vs. Mock βIII tubulin: miR‐326 P = 0.0027; miR‐326 and ARRB1 P = 0.046, GFAP: miR‐326 P = 0.0002; miR‐326 and ARRB1 P = 0.011). Data represent means ± SD from five independent experiments. Statistics: One‐way ANOVA and two‐way ANOVA, * P < 0.05; **P < 0.01; *** P < 0.001; **** P < 0.0001 vs. indicated controls.

Article Snippet: Cell was stained with mouse monoclonal antibodies against βIII tubulin (MAB 1637; Merck, Darmstadt, Germany) and GFAP (MAB360).

Techniques: Expressing, Transfection, MTT Assay, Quantitative RT-PCR

Journal: Cells

Article Title: Prolonged Heat-Treated Mesenchymal Precursor Cells Induce Positive Outcomes Following Transplantation in Cervical Spinal Cord Injury

doi: 10.3390/cells14191488

Figure Lengend Snippet: Axonal and glia profile 8 weeks post-injury. Images showing a stitched section of the spinal cord with the injury site labeled with βIII-tubulin+ axonal profile in mice receiving ( A ) HBSS, ( B ) dMPC and ( C ) MPC treatments. The corresponding ( ii ) images show the microscopic view of the injury site as outlined by the white boxes. Images showing a stitched section of the spinal cord with the injury site labeled with GFAP+ astrocytic glia profiles in mice receiving ( D ) HBSS, ( E ) dMPC and ( F ) MPC treatment. The corresponding ( ii ) images show the microscopic view of the injury site as outlined by the white boxes; * shows areas devoid of axons. Scale bar = 200 μm.

Article Snippet: For the dual-injection study, the primary antibodies used were βIII tubulin (mouse, 1:800; Covance, Princeton, NJ, USA), GFAP, anti-APC (mouse, 1:200; Abcam, Cambridge, MA, USA) and green fluorescence protein (GFP, chicken, 1:10,000; Aves Lab, Davis, CA, USA).

Techniques: Labeling

Journal: Cells

Article Title: Prolonged Heat-Treated Mesenchymal Precursor Cells Induce Positive Outcomes Following Transplantation in Cervical Spinal Cord Injury

doi: 10.3390/cells14191488

Figure Lengend Snippet: GFP+ NSCs in spinal cord sections of dMPC_D1/NSC_D3 mice. Gross overview of surviving GFP+ NSCs transplanted at day 3 after intravenous injection of dead MPCs at D1 following cervical spinal cord injury showing ( Ai – Aiii ) βIII-tubulin and GFP. High-magnification images as outlined in the white boxes are shown in ( Aiv – Aviii ) as follows: ( Aiv ) GFP+ astrocytes with βIII-tubulin, ( Av ) βIII-tubulin+ axonal profiles, ( Avi ) GFP+ astrocytes, ( Avii ) GFP+ oligodendrocytes with βIII-tubulin and ( Aviii ) GFP+ oligodendrocytes. The white arrows seen in ( Aiv , Avi ) show an astrocyte projecting long processes. Astrocytic profile of the spinal cord tissue around the injury site is seen in ( Bi – Biii ) showing GFAP+ astrocytes and GFP+ differentiated NSCs. ( Biv , Bv ) shows high-magnification image of the area outline in the white boxes in ( Bi ). The orange lines intersecting the images show the dual label of GFAP and GFP on the cell that the line intersects with in the vertical column on the right and horizontal column underneath. The blue dotted boxes in ( Bi ) correspond to the figures in ( Ci , Cii ) showing the oligodendrocyte profile of the spinal cord tissue around the injury site with APC+ oligodendrocytes and GFP+ differentiated NSCs. The high-magnification image of the area in the white box in ( Cii ) is shown in ( Ciii ). ( Civ ) shows the GFP+-labeled oligodendrocyte outlined by the white box in ( Ciii ). ( Cv ) is the 3D view of the same cell as shown by the pink arrow. Scale bars for ( Ai ), ( Bi ), ( Ci ), ( Cii ) = 200 μm, ( Ciii ) = 50 μm and ( Aiv ), ( Avii ), ( Biv ), ( Bv ), ( Civ ), ( Cv ) = 10 μm.

Article Snippet: For the dual-injection study, the primary antibodies used were βIII tubulin (mouse, 1:800; Covance, Princeton, NJ, USA), GFAP, anti-APC (mouse, 1:200; Abcam, Cambridge, MA, USA) and green fluorescence protein (GFP, chicken, 1:10,000; Aves Lab, Davis, CA, USA).

Techniques: Injection, Labeling

Journal: Cells

Article Title: Prolonged Heat-Treated Mesenchymal Precursor Cells Induce Positive Outcomes Following Transplantation in Cervical Spinal Cord Injury

doi: 10.3390/cells14191488

Figure Lengend Snippet: GFP+ NSCs in spinal cord sections of dMPC_D1/NSC_D7 mice. Gross overview of surviving GFP+ NSCs transplanted at day 7 after intravenous injection of dead MPCs at D1 following cervical spinal cord injury showing ( Ai – Aiii ) βIII-tubulin and GFP. High-magnification images as outlined in the white boxes are shown in iv-viii as follows: ( Aiv ) GFP+ astrocytes with βIII-tubulin, ( Av ) βIII-tubulin+ axonal profiles, ( Avi ) GFP+ astrocytes, ( Avii ) GFP+ oligodendrocytes with βIII-tubulin, ( Aviii ) βIII-tubulin+ axonal profiles and ( Aix ) GFP+ oligodendrocytes. The white arrows seen in ( Aiv ) show astrocytes projecting long processes. Astrocytic profile of the spinal cord tissue around the injury site is seen in ( Bi – Biii ) showing GFAP+ astrocytes and GFP+ differentiated NSCs. ( Biv ) shows a high-magnification image of the area outline in the white box in ( Bi ). ( Bv , Bvi ) shows the 3D view of the same image in ( Biv ) showing the dual label of cells with GFAP and GFP. ( Bvii ) shows the dual-labeled high-magnification image of the area outline in the white box in Bi, while the single-channel image is seen in ( Bviii ) for the GFAP+ axonal profile and ( Bix ) for GFP+ oligodendrocytes. Oligodendrocyte profile of the spinal cord tissue around the injury site is seen in ( Ci – Ciii ) showing APC+ oligodendrocytes and GFP+ differentiated NSCs. ( Civ ) shows the dual-labeled high-magnification image of the area outline in the white box in ( Ci ), while ( Cv ) shows the 3D view of the same area. ( Cvi ) shows the dual-labeled high-magnification images of the area outline in the white box in ( Ci ), while ( Cvii , Cviii ) show the 3D view of the same area. Scale bars for ( Ai ), ( Bi ), ( Ci ), ( Cii ) = 200 μm, while the rest of the scale bars = 10 μm.

Article Snippet: For the dual-injection study, the primary antibodies used were βIII tubulin (mouse, 1:800; Covance, Princeton, NJ, USA), GFAP, anti-APC (mouse, 1:200; Abcam, Cambridge, MA, USA) and green fluorescence protein (GFP, chicken, 1:10,000; Aves Lab, Davis, CA, USA).

Techniques: Injection, Labeling

Journal: Cells

Article Title: Prolonged Heat-Treated Mesenchymal Precursor Cells Induce Positive Outcomes Following Transplantation in Cervical Spinal Cord Injury

doi: 10.3390/cells14191488

Figure Lengend Snippet: Intraspinal-injected NSCs differentiated into astrocytes with long processes. High-magnification image of an area around injection site (*) in a mouse receiving dMPC_D1/NSC_D7 showing differentiated NSCs into astrocytes projecting long processes with ( A ) dual label of βIII-tubulin and GFP, ( B ) for single-channel view of βIII-tubulin and ( C ) single-channel view of GFP. Scale bar = 10 μm.

Article Snippet: For the dual-injection study, the primary antibodies used were βIII tubulin (mouse, 1:800; Covance, Princeton, NJ, USA), GFAP, anti-APC (mouse, 1:200; Abcam, Cambridge, MA, USA) and green fluorescence protein (GFP, chicken, 1:10,000; Aves Lab, Davis, CA, USA).

Techniques: Injection

Journal: PLoS ONE

Article Title: Interferon Gamma and Sonic Hedgehog Signaling Are Required to Dysregulate Murine Neural Stem/Precursor Cells

doi: 10.1371/journal.pone.0043338

Figure Lengend Snippet: γ in NSPCs are blocked by cyclopamine. In Figure a: Photomicrographs of undifferentiated murine NSPCs are given. The left panel shows an immunocytochemical staining for GFAP, the middle panel for βIII-tubulin and the right panel shows both. The first row represents the control, and the second row represents the IFNγ-treated cells. The third row shows cells that are treated with cyclopamine alone, and the fourth row shows cells treated with both substances. In Figure b: The results of the real-time quantitative PCR experiments for GFAP, βIII-tubulin, Stat 1 and SHH gene expression are depicted. Delta CT values are means +/− standard error of mean (SEM). Independent experiments were repeated for three times in triplicate.

Article Snippet: Primary antibodies used at 4°C overnight were the monoclonal mouse antibody against βIII-tubulin (Tuj1; 1∶500; R&D Systems, Minneapolis, USA or 1∶800, Abcam, Cambridge, UK) and the polyclonal rabbit antibody against glial fibrillaric acid protein (GFAP) (1∶500; Dako, Hamburg, Germany or 1∶1000; Abcam).

Techniques: Staining, Control, Real-time Polymerase Chain Reaction, Gene Expression