Journal: Stem cells (Dayton, Ohio)

Article Title: Mifepristone-inducible caspase-1 expression in mouse embryonic stem cells eliminates tumor formation but spares differentiated cells in vitro and in vivo.

doi: 10.1002/stem.1000

Figure Lengend Snippet: Figure 4. Survival of differentiated dopamine neurons derived from caspase-1-ESCs treated with mifepristone in vitro. (A): Mifepristone had no effect on differentiated caspase-1-ESCs after 6 days treatment. Scale bar ¼ 500 lm. (B): Mifepristone had no effect on differentiated WT and cas- pase-1-ESCs. (C): Fluorescence-activated cell sorting (FACS) results showed that mifepristone did not affect differentiated WT or caspase-1-ESCs; (C0) was the quantitative analysis from (C) (n ¼ 3). (D): Mifepristone did not induce DNA cleavage in differentiated caspase-1-ESCs. Scale bar ¼ 10 lm. (E): The protein levels of caspase-1, p20 and p10 expression in differentiated caspase-1-ESCs showed increase after mifepristone treatment; (E0) was the quantitative analysis from (E) (n ¼ 3). *, p < .05 (compared with caspase-1-ES vehicle). (F): MAP2þ/THþ caspase-1-ESCs survived in the presence of mifepristone. Scale bar ¼ 50 lm. (G): Mifepristone treatment did not affect THþ or MAP2þ colony proportion. Colony counts were per- formed in triplicate and at least 200 colonies were counted each time. Abbreviations: ESC, embryonic stem cell; MAP2, microtubule-associated pro- tein 2; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PI, propidium iodide; TH, tyrosine hydroxylase; WT, wild type

Article Snippet: After fixing the cells and sections, primary antibodies were applied as follows: mouse anti–microtubule-associated protein 2 (anti-MAP2) antibody (1:2,000, M4403, Sigma), rabbit anti-tyrosine hydroxylase (TH) antibody (1:500, AB152, Chemicon), mouse anti–sex-determining region Y protein (anti-SRY) antibody (1:200, ab22166, Abcam, UK, www.abcam.com), rabbit anti-Oct-4 antibody (1:200, 2750, Cell Signaling), mouse anti-nestin antibody (1:200, MAB353, Chemicon), and goat anti-MAP2 antibody (1:20, sc-5359, Santa Cruz).

Techniques: Derivative Assay, In Vitro, Fluorescence, FACS, Expressing

Journal: Stem cells (Dayton, Ohio)

Article Title: Mifepristone-inducible caspase-1 expression in mouse embryonic stem cells eliminates tumor formation but spares differentiated cells in vitro and in vivo.

doi: 10.1002/stem.1000

Figure Lengend Snippet: Figure 5. Ablation of tumors and survival of dopamine neurons derived from caspase-1-ESCs in brain after mifepristone treatment. (A): Undifferenti- ated WT ESCs and caspase-1-ESCs were Oct-4þ. Most differentiated WT and caspase-1-ESCs were nestinþ. Scale bar ¼ 10 lm. (B): The mifepristone treatment increased the life span of mice transplanted with undifferentiated caspase-1-ESCs (n ¼ 6). (C): WT ESCs formed tumors in the absence or pres- ence of mifepristone, while caspase-1-ESCs formed tumors only in the absence of mifepristone. (D): The hematoxylin and eosin staining verified that the tumors were derived from ESCs. Scale bar ¼ 50 lm. (E): Dopamine neurons derived from caspase-1-ESCs appeared in both vehicle- and mifepristone- treated mice brains. The arrows indicated the SRYþ/THþ/MAP2þ cells. (F): Quantitative analysis of SRYþ/THþ/MAP2þ cells in the brain showed that mifepristone did not affect dopamine neurons derived from caspase-1-ESCs (n ¼ 4). Scale bar ¼ 10 lm. Abbreviations: ESC, embryonic stem cell; MAP2, microtubule-associated protein 2; SRY, sex-determining region Y protein; TH, tyrosine hydroxylase; WT, wild type.

Article Snippet: After fixing the cells and sections, primary antibodies were applied as follows: mouse anti–microtubule-associated protein 2 (anti-MAP2) antibody (1:2,000, M4403, Sigma), rabbit anti-tyrosine hydroxylase (TH) antibody (1:500, AB152, Chemicon), mouse anti–sex-determining region Y protein (anti-SRY) antibody (1:200, ab22166, Abcam, UK, www.abcam.com), rabbit anti-Oct-4 antibody (1:200, 2750, Cell Signaling), mouse anti-nestin antibody (1:200, MAB353, Chemicon), and goat anti-MAP2 antibody (1:20, sc-5359, Santa Cruz).

Techniques: Derivative Assay, Staining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Expression of the Interleukin 6 System in Cortical Lesions From Patients With Tuberous Sclerosis Complex and Focal Cortical Dysplasia Type IIb

doi: 10.1097/nen.0b013e3181eaeae5

Figure Lengend Snippet: FIGURE 2. Cell-specific distribution of IL-6 in apparently normal CTX, TSC tubers and FCDIIb lesions. (A, B) Hematoxylin and eosin (H&E) staining. Representative photomicrographs of TSC (A) and FCDIIb (B) showing areas of cortical disorganization containing different cell types, including DNs (arrowheads) with high Nissl substance staining and diverse directionality, TS cells with eosinophilic cytoplasm (arrows in A), and BCs with opalescent cytoplasm and eccentric nuclei (arrows in B). (C, D) Weak to moderate IL-6 IR in neurons (arrows in C), endothelial cells (double arrows in D), and glia-like cells (arrowheads in C and D) in neocortex and white matter (WM). (EYG) Strong IL-6 IR in TS cells (insert, E, arrow) and in DNs of different size and shape (arrowheads in EYG). Merged images show colocalization of IL-6 (green) with NeuN (red) (insert, F), and colocalization of IL-6 (green) with MAP2 (red) (insert, G) in DNs. (H) Representative confocal image showing no IL-6 IR (green) colocalization with GFAP (red). (I and J) Interleukin 6 IR in FCDIIb. Weak (double arrows in I) to moderate (arrows and in I insert) IL-6 IR in BCs; strong IL-6 IR in DNs (arrowhead, I) and in GNs with central nuclei (J). Merged images show colocalization of IL-6 (green) with NeuN (red) in GNs (insert, J) and of IL-6 (green) with NF200 (red) in DNs (K). (L) Confocal image shows that no IL-6 IR DNs (arrowhead) or GNs (double arrowheads) are colocalized with GFAP (red, arrows). Merged image show most cells with IL-6 IR (green, BCs) colabeled with GFAP (red) (insert in L). Sections are counterstained with hematoxylin (CYG, I, J) or Hoechst 33258 (inserts in G, K, L). Bars = (AYE, H, L) 50 Km; (F, G, J, K) 10 Km; (I) 25 Km.

Article Snippet: Unauthorized reproduction of this article is prohibited. and antiYglial fibrillary acidic protein (GFAP, mouse monoclonal, 1:1000; Sigma), ii) antiYIL-6 (1:100) or antiYIL-6R (rabbit polyclonal, 1:100) and anti-NeuN (mouse monoclonal, 1:500; Chemicon), iii) antiYIL-6 (1:100) and anti-microtubuleassociated protein 2 (MAP2) (mouse monoclonal, 1:100; Boster), and iv) antiYIL-6 (rabbit polyclonal, 1:100) or antiYIL-6R (rabbit polyclonal, 1:100) and anti-NF200 (mouse monoclonal, 1:100; Boster) in 0.01 M PBS (pH 7.4) containing 1% bovine serum albumin overnight at 4-C. After 3 rinses, the sections were incubated with a mixture of goat anti-rabbit immunoglobulin class GYfluorescein isothiocyanate (1:200; Zhongshan Goldenbridge Biotechnology Co, Beijing, China) and goat antimouse immunoglobulin class GYtetramethyl rhodamine isothiocyanate (1:200; Upstate Biotechnology, Lake Placid, NY) for 1 hour at 37-C. Counterstaining of cell nuclei was performed by incubating the sections with Hoechst 33258 (0.1 Kg/mL; Sigma).

Techniques: Staining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Expression of the Interleukin 6 System in Cortical Lesions From Patients With Tuberous Sclerosis Complex and Focal Cortical Dysplasia Type IIb

doi: 10.1097/nen.0b013e3181eaeae5

Figure Lengend Snippet: FIGURE 3. Cell-specific distribution of IL-6Rs in apparently normal CTX, TSC tubers, and FCDIIb cortical lesions. (A) No IL-6R IR is seen in neuronal or glial cells in control CTX. (BYH) Tuberous sclerosis complex. Strong IL-6R IR in DNs of different sizes and shapes in the neocortex and white matter (WM) (arrowheads in BYD) and in TSC cells (insert in D, arrow). Merged images show colocalization of IL-6R (green) with MAP2 (red) (insert in B), of IL-6R (green) with NeuN (red) (insert in C), and of IL-6R (green) with NF200 (red) (EYG) in DNs (arrowheads) and TS cells (arrows), respectively. (H) Representative confocal image showing that no IL-6R+ (green) DNs (arrowheads) or TS cells (arrows) are colocalized with GFAP (red). Scattered GFAP+ reactive astrocytes are colabeled with IL-6R (insert in H, indicated by double arrows). (IYL) Interleukin 6 receptor in FCDIIb. Moderate to strong IL-6R IR in BCs (I, arrows and double arrows). Interleukin 6 receptorYnegative BCs are indicated by an asterisk (J). Strong IL-6R IR was also observed in GNs (K) and DNs (L). Merged images show colocalization of IL-6R (green) with NeuN (red) in GNs (arrowhead) and BCs (arrow) (insert in I) and colocalization of IL-6R (green) with NeuN (red) in DNs (insert in L), respectively. (M) Representative confocal image showing that no IL-6R-positive (green) GNs or DNs (arrowheads) are colocalized with GFAP (red). Merged image showing that most IL-6R+ (green) BCs colabeled with GFAP (red) (arrows in M). Interleukin 6 receptorYnegative BCs are indicated by double arrows. Sections are counterstained with hematoxylin (BYD, IYL) and Hoechst 33258 (insert in B, C; G, H, M). Scale bar = (A) 100 Km; (B, EYG) 30 Km; (C, D, J, L) 20 Km; (H, I, M) 50 Km; (K) 15 Km.

Article Snippet: Unauthorized reproduction of this article is prohibited. and antiYglial fibrillary acidic protein (GFAP, mouse monoclonal, 1:1000; Sigma), ii) antiYIL-6 (1:100) or antiYIL-6R (rabbit polyclonal, 1:100) and anti-NeuN (mouse monoclonal, 1:500; Chemicon), iii) antiYIL-6 (1:100) and anti-microtubuleassociated protein 2 (MAP2) (mouse monoclonal, 1:100; Boster), and iv) antiYIL-6 (rabbit polyclonal, 1:100) or antiYIL-6R (rabbit polyclonal, 1:100) and anti-NF200 (mouse monoclonal, 1:100; Boster) in 0.01 M PBS (pH 7.4) containing 1% bovine serum albumin overnight at 4-C. After 3 rinses, the sections were incubated with a mixture of goat anti-rabbit immunoglobulin class GYfluorescein isothiocyanate (1:200; Zhongshan Goldenbridge Biotechnology Co, Beijing, China) and goat antimouse immunoglobulin class GYtetramethyl rhodamine isothiocyanate (1:200; Upstate Biotechnology, Lake Placid, NY) for 1 hour at 37-C. Counterstaining of cell nuclei was performed by incubating the sections with Hoechst 33258 (0.1 Kg/mL; Sigma).

Techniques: Control

Journal: Current Issues in Molecular Biology

Article Title: Hydrogen Peroxide Induced Toxicity Is Reversed by the Macrocyclic IRAP-Inhibitor HA08 in Primary Hippocampal Cell Cultures

doi: 10.3390/cimb44100340

Figure Lengend Snippet: Characterisation of primary hippocampal cell culture. Representative images of ( A ) the expression of microtubule-associated protein 2 (MAP2) shown in green, ( B ) the expression of glial fibrillary acidic protein (GFAP) shown in red and ( C ) the cell nuclei stained with 4′,6-diamidino-2-phenylindole (DAPI) shown in blue in a primary hippocampal culture. ( D ) MAP2 (green), GFAP (red) and DAPI (blue) are merged. The images were acquired using ImageXpress (Molecular Devices) mounted with a 20× objective.

Article Snippet: The antibodies mouse anti-MAP2 (Merck Millipore, Burlington, MA, USA) and rat anti-GFAP (Invitrogen, Waltham, MA, USA) were added in concentration 1:500 for 1 h in RT.

Techniques: Cell Culture, Expressing, Staining

Journal: Chinese Medical Journal

Article Title: Automated quantification of Ki-67 index associates with pathologic grade of pulmonary neuroendocrine tumors

doi: 10.1097/cm9.0000000000000109

Figure Lengend Snippet: Figure 1: (A) Typical carcinoid, with trabecular pattern with delicate intervening vascular stroma, no mitoses or necrosis are identified; (B) Ki-67, showing 12.66% positivity by computer image analysis and 15% positivity by manual counting in the hot spot; (C–E). TC showed diffuse positive expression for CD56, synaptophysin, CgA and MAP2, respectively (A: Hematoxylin-eosin staining; B–E: Immunohistochemistry staining. The original magnification of A, B was 100, magnification for remaining cases were 200).

Article Snippet: The following neuroendocrine and proliferative immunohistochemical markers were used: monoclonal mouse anti-human synaptophysin (clone DAK-SYNAP; Dako, Glostrup, Denmark), monoclonal rabbit anti-human chromogranin A (clone EP38; Zhongshan Golden Bridge Biotechnology, Beijing, China), monoclonal mouse anti-human CD56 (Clone 123C3; Dako), monoclonal mouse anti-human MAP2 (Clone AP18; Zhongshan Golden Bridge Biotechnology), and monoclonal mouse anti-human Ki-67 (clone MIB-1; Dako).

Techniques: Expressing, Staining, Immunohistochemistry