Journal: International Journal of Molecular Sciences

Article Title: Development of a High-Efficacy Reprogramming Method for Generating Human Induced Pluripotent Stem (iPS) Cells from Pathologic and Senescent Somatic Cells

doi: 10.3390/ijms21186764

Figure Lengend Snippet: TRA-1-60 and alkaline phosphatase staining of iPS cell colonies induced from the dermal tissue of three elderly patients. ( a ) TRA-1 immunofluorescent staining of iPS cell colonies. Immunofluorescent staining with anti-TRA1-60 antibody was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 35 post-transfection. An integration of 81 continuous fields of the view captured with the Keyence BZ-X710 microscope is shown (observed area: 23.915 mm × 17.936 mm). BF, bright field. Arrows indicate TRA1-60-positive iPS cell colonies. Scale bars: 3.0 mm. ( b ) Alkaline phosphatase staining of iPS cell colonies. Staining with alkaline phosphatase was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 36 post-transfection. Arrows indicate alkaline phosphatase-positive iPS cell colonies. ( c ) The number of iPS cell colonies stained by alkaline phosphatase was quantitively analyzed in all fibroblasts. The left side is protocol 1 and the right side is protocol 2. Values shown are the mean ± SEM (* p < 0.05, n = 3).

Article Snippet: An Anti-TRA-1-60 antibody, anti-SSEA4 antibody, and alkaline phosphatase stain kit (AP Staining Kit, Red-Color) was purchased from Funakoshi ® (Tokyo, Japan).

Techniques: Staining, Transfection, Microscopy

Journal:

Article Title: Characterization of Monoclonal Antibodies Raised against the Latent Nuclear Antigen of Human Herpesvirus 8

doi:

Figure Lengend Snippet: LN53 reactivity to LNA in nodular KS. Immunocytochemistry was performed on a paraffin-embedded tumor from classical KS. Permeabilization was performed by microwave. Negative control biopsies were from angiosarcomas and hemangiomas (not shown). After incubation with normal rabbit serum (DAKO), MAbs were applied for 1 h at 22°C followed by two washes in PBS–0.1% (vol/vol) Tween. The secondary biotin-conjugated antibody (rabbit anti-rat; DAKO) was applied for 30 min followed by washing. Antibody reactions were visualized with streptavidine-alkaline phosphatase (Vector Laboratories) and a substrate red chromogen (Vector). Adjacent sections were stained by standard methods. (a) Hematoxylin and eosin staining showing KS nodule (left) and surrounding dermis (right). (b) Adjacent sections stained with LN53 showing clear demarcation between LNA expression in the KS lesion and not in the surrounding dermis (magnification, ×40). Almost all spindle-shaped cells are positive for LNA (×60) (c), with a stippling pattern reminiscent of the staining of PEL cells (×160) (d).

Article Snippet: Antibody reactions were visualized with streptavidine-alkaline phosphatase (Vector Laboratories) and a substrate red chromogen (Vector).

Techniques: Immunocytochemistry, Negative Control, Incubation, Plasmid Preparation, Staining, Expressing