Journal: Journal of Clinical Microbiology

Article Title: Human Immunodeficiency Virus Type 1 Protease Inhibitor Drug-Resistant Mutants Give Discordant Results When Compared in Single-Cycle and Multiple-Cycle Fitness Assays ▿

doi: 10.1128/JCM.00605-10

Figure Lengend Snippet: HIV-1 vectors pDAT1, used for the SCA (A), and pAT1, used for the MCA (B). The open reading frames for gag, pol, the accessory genes, Thy1 markers, and long terminal repeats are shown, as are the bacterial pUC19 sequences (pUC) and the 5′ cellular flanking sequence. Vertical lines indicate the locations of restriction enzymes. A black triangle indicates the location of the deletion in the envelope in the pDAT vector used for the SCA.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated-mouse anti-rat Thy1.1 (HIS51) and R-phycoerythrin (PE)-conjugated rat anti-mouse Thy1.2 (30-H12) monoclonal antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Sequencing, Plasmid Preparation

Journal: Journal of Clinical Microbiology

Article Title: Human Immunodeficiency Virus Type 1 Protease Inhibitor Drug-Resistant Mutants Give Discordant Results When Compared in Single-Cycle and Multiple-Cycle Fitness Assays ▿

doi: 10.1128/JCM.00605-10

Figure Lengend Snippet: Experimental design of the SCA. Circular versions of the constructs from Fig. ​Fig.11 are shown with the wild type in black and the mutant in gray. COS-1 or 293 cells were cotransfected with pHCMV-g or pSV-A-MLV-ENV and either wild-type pDAT1 (pDAT1WT) or mutant pDAT2 (pDAT2MUT). Wild-type (black) and mutant (gray) virus stocks are represented by hexagons. The titers of virus stocks were determined by measuring the p24 or VSV-g protein content. PM1 cells were coinfected with equal amounts of wild-type and mutant viruses. After 40 h, cells were harvested, and a subset of cells was stained with either an anti-Thy1.1 antibody, an anti-Thy1.2 antibody, or both. Four different types of cells can be observed during analysis: uninfected cells, cells infected with wild-type virus only, cells infected with mutant virus only, and cells infected with both viruses (dually infected).

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated-mouse anti-rat Thy1.1 (HIS51) and R-phycoerythrin (PE)-conjugated rat anti-mouse Thy1.2 (30-H12) monoclonal antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Construct, Mutagenesis, Staining, Infection

Journal: Biomolecules

Article Title: Mitochondria Transfer from Adipose Stem Cells Improves the Developmental Potential of Cryopreserved Oocytes

doi: 10.3390/biom12071008

Figure Lengend Snippet: Mouse adipose stem cell (ASC) isolation and confirmation. ( A ). Morphology of ASC shows regular fibroblastic cell morphology. ( B ). Flow cytometric analysis of cultured ASC at 3rd passage. ASC was positive for CD29, CD90, and CD105 and negative for CD45, which confirms the purity of ASC. ( C ). Immunofluorescence staining confirmed the expression of CD29 and CD105 in ASCs. The nucleus was stained with DAPI. ( D , E ). Confirmation of the multipotency of ASC. ( D ). Adipogenic differentiation. Oblate-like cell morphology and numerous lipid droplets can be visualized without specific staining. E. Osteogenic differentiation. The cells developed a cluster-shaped structure and showed positive red staining with alizarin red staining, confirming the multipotency of ASC.

Article Snippet: The cells were then incubated with Alexa fluor 647 anti-mouse CD29 (Biolegend, Tokyo, Japan), Alexa fluor 488 anti-mouse CD105 (Biolegend), phycoerythrin anti-mouse CD90 (Novus Biologicals, Littleton, CO, USA), FITC anti-mouse CD45 (Invitrogen, Waltham, MA, USA) fluorescent conjugated antibodies or isotype control Alexa flour 647 anti-mouse IgG1(Biolegend), and FITC anti-mouse IgG1 (Biolegend) antibodies according to recommended concentration and incubated for 1 h at RT.

Techniques: Isolation, Cell Culture, Immunofluorescence, Staining, Expressing