Journal: Journal of Alzheimer's disease : JAD

Article Title: Intraneuronal Amylin Deposition Peroxidative Membrane Injury and Increased IL-1β Synthesis in Brains of Alzheimer’s Disease Patients with Type-2 Diabetes and in Diabetic HIP Rats

doi: 10.3233/JAD-160047

Figure Lengend Snippet: Amylin-mediated peroxidative membrane injury increases IL-1β synthesis in cultured neurons. A) Lipid peroxidation measurements with C11-BODIPY581/591 in cultured primary hippocampal rat neurons incubated for two hours under control conditions (Ctl), with aggregated amylin (Amy), with 50 μM poloxamer 188 followed by aggregated amylin (S→Amy), and with 5 mM NAC for 30 min followed by aggregated amylin (NAC→Amy). B) IL-1β level, assessed by immunofluorescence, in cultured primary hippocampal rat neurons incubated under the four conditions described in panel A. Neuronal cells were identified with mouse anti-MAP2 antibody. 10 neurons/rat; n=5 rats/group. Data are presented as mean ± standard error. **p < 0.01, ***p < 0.001.

Article Snippet: Primary neuronal cells were detected with mouse anti-MAP2 antibody (NB300-213; Novus Biologicals; CO).

Techniques: Membrane, Cell Culture, Incubation, Control, Immunofluorescence

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