Journal: Cell reports

Article Title: Methylation of dual-specificity phosphatase 4 controls cell differentiation

doi: 10.1016/j.celrep.2021.109421

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ImmPACT Vector Red Substrate Kit , Vector Laboratories , Cat# SK-5105; RRID:AB_2336524.

Techniques: Plasmid Preparation, Virus, Recombinant, Protease Inhibitor, Modification, Protein Purification, Western Blot, Membrane, Viability Assay, Polymer, cDNA Synthesis, Sequencing, Real-time Polymerase Chain Reaction, Software

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: Cell Tropism of Simian Immunodeficiency Virus in Culture Is Not Predictive of in Vivo Tropism or Pathogenesis

doi:

Figure Lengend Snippet: SIV in Situ Hybridization Results

Article Snippet: If immunofluorescence followed the in situ hybridization, 2-hydroxy-3-naphtoic acid-2′-phenylaniide phosphate (HNPP) fluorescent detection system (Boehringer Mannheim) was used.

Techniques: In Situ Hybridization

Journal:

Article Title: Cell Tropism of Simian Immunodeficiency Virus in Culture Is Not Predictive of in Vivo Tropism or Pathogenesis

doi:

Figure Lengend Snippet: Localization of virus in macaques infected with SIVmac239/316. A: SIV in situ hybridization reveals a dendritic staining pattern at 21 days after infection in lymph node consistent with antigen/antibody trapping on follicular dendritic cells. B: Numerous in situ hybridization-positive cells at 50 days after infection in lymph node. The higher magnification inset illustrates the cell morphology. C: In situ hybridization for SIV (blue) followed by immunohistochemistry for the macrophage marker CD68 (red) in LN at 50 days after infection. No co-localization of the two labels (would appear purple) is evident.

Article Snippet: If immunofluorescence followed the in situ hybridization, 2-hydroxy-3-naphtoic acid-2′-phenylaniide phosphate (HNPP) fluorescent detection system (Boehringer Mannheim) was used.

Techniques: Virus, Infection, In Situ Hybridization, Staining, Immunohistochemistry, Marker

Journal:

Article Title: Cell Tropism of Simian Immunodeficiency Virus in Culture Is Not Predictive of in Vivo Tropism or Pathogenesis

doi:

Figure Lengend Snippet: Immunophenotype of SIV-infected cells in SIVmac239/316 infection. A: Double-label confocal microscopy (three channel) in LN at 50 days after infection. Images for individual channels (CD3 with Alexa 488, green; SIV in situ hybridization with Fast Red, red, and differential interference contrast, DIC) are shown on the left and a large merged image containing two channels plus DIC showing many double-positive cells on the right. At higher magnification (inset with scale bar) the infected cells can be seen to be morphologically consistent with lymphocytes. B: In lymph node in long-term infected animals, images for individual channels (SIV-ISH, red; Ham56-macrophages, green; and DIC) are shown on the left. No evidence of macrophage infection (co-localization of the two markers) is present in the larger merged image. C: Infection of macrophages in vivo is extremely rare in SIVmac239/316 infection. C shows the only instance in which macrophage infection could be demonstrated (CD68 with Alexa 488, green; SIV-ISH, red; and DIC). The double-positive cell is present in the subcapsular sinus of the axillary lymph node. D: Co-localization of SIV (red), macrophages (HAM56, green), and T cells (CD5, blue) in the lymph node of a chronically infected animal. In the larger merged image several SIV-infected T cells are present and appear pink/purple because of the mixing of red and blue labels.

Article Snippet: If immunofluorescence followed the in situ hybridization, 2-hydroxy-3-naphtoic acid-2′-phenylaniide phosphate (HNPP) fluorescent detection system (Boehringer Mannheim) was used.

Techniques: Infection, Confocal Microscopy, In Situ Hybridization, In Vivo

Journal:

Article Title: Cell Tropism of Simian Immunodeficiency Virus in Culture Is Not Predictive of in Vivo Tropism or Pathogenesis

doi:

Figure Lengend Snippet: Total numbers of SIV-infected cells/mm2 (left axis) versus numbers of SIV-infected macrophages/mm2 (right axis) detected by combined RNA in situ hybridization/immunohistochemistry for SIV and macrophages using HAM56 in LN and spleen. Bars represent averages of five 1-mm2 areas per slide from each tissue in two animals per time point. In SIVmac239/316-infected animals both the total numbers of infected cells and infected macrophages were lower than animals inoculated with SIVmac239. All animals infected with SIVmac239 for 21 days or more had evidence of SIV-infected macrophages. In terminal SIVmac239-infected animals, the number of macrophages was primarily represented by multinucleated giant cells. These cells were not observed in terminal SIVmac239/316-infected animals.

Article Snippet: If immunofluorescence followed the in situ hybridization, 2-hydroxy-3-naphtoic acid-2′-phenylaniide phosphate (HNPP) fluorescent detection system (Boehringer Mannheim) was used.

Techniques: Infection, RNA In Situ Hybridization, Immunohistochemistry