Journal: Regenerative Therapy

Article Title: Multilineage-differentiating stress-enduring (Muse)-like cells exist in synovial tissue

doi: 10.1016/j.reth.2018.10.005

Figure Lengend Snippet: Expression of stemness markers by M- and SY-clusters. Immunostaining of clusters formed from Muse-like cells and unsorted synovial cells. Clusters were positive for NANOG, OCT3/4, and SOX2. Scale bars = 50 μm. DAPI was used as a counterstain.

Article Snippet: The blocking solution was discarded, and the slides were incubated with the following primary antibodies: polyclonal rabbit anti-NANOG antibody (1:500; Merck Millipore), polyclonal rabbit anti-OCT3/4 antibody (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), and polyclonal rabbit anti-SOX2 antibody (1:500; Abcam, Cambridge, UK) in antibody diluent (1% normal goat serum and 0.1% Triton X-100 in 0.02 M PBS) at 4 °C overnight.

Techniques: Expressing, Immunostaining

Journal: PLoS ONE

Article Title: Expression of Pluripotency Master Regulators during Two Key Developmental Transitions: EGA and Early Lineage Specification in the Bovine Embryo

doi: 10.1371/journal.pone.0034110

Figure Lengend Snippet: Primers used for the qRT-PCR of different genes.

Article Snippet: NANOG was labelled with a rabbit polyclonal anti-NANOG antibody (Reprotech; 1∶500 in PBS–BSA for immunofluorescence and 1/1000 in PBS–T 4% non-fat dried milk for Western blot).

Techniques:

Journal: PLoS ONE

Article Title: Expression of Pluripotency Master Regulators during Two Key Developmental Transitions: EGA and Early Lineage Specification in the Bovine Embryo

doi: 10.1371/journal.pone.0034110

Figure Lengend Snippet: Expression profiles of OCT4 , SOX2 and NANOG ( A–C ) were charted using qRT-PCR at pre-implantation stages of bovine oocytes/embryos: GV (Germinal Vesicle), MII (Metaphase 2 oocyte), 4-cell stage, 8–16-cell stage, Morula (25–30 cells) and E7 (Day 7 blastocyst). The results were normalised using the geometric mean of the endogenous GAPD, SDHA and YWHAZ genes. Values are means ± SEM of one equivalent embryo/oocyte. The experiment was repeated three times and each repetition contained a triplicate of each sample stage. The localization and compartmentalization of OCT4 (D–F), SOX2 (G–I) and NANOG (J–L) was determined by whole mount in situ hybridization (ISH) in a spatio-temporal manner from day7 to day9 (E7, E8 and E9) blastocysts. Encircled regions demarcate the ICM.

Article Snippet: NANOG was labelled with a rabbit polyclonal anti-NANOG antibody (Reprotech; 1∶500 in PBS–BSA for immunofluorescence and 1/1000 in PBS–T 4% non-fat dried milk for Western blot).

Techniques: Expressing, Quantitative RT-PCR, In Situ Hybridization

Journal: PLoS ONE

Article Title: Expression of Pluripotency Master Regulators during Two Key Developmental Transitions: EGA and Early Lineage Specification in the Bovine Embryo

doi: 10.1371/journal.pone.0034110

Figure Lengend Snippet: Immunofluorescent detection of OCT4, SOX2 and NANOG protein was done in the pre-implantation bovine embryos/oocytes: GV (Germinal Vesicle), MII (Metaphase 2 oocyte), 4-cell stage, 8–16-cell stage and Morula (25–30 cells) using immunofluorescence (A) and Western Blot (B-D). Protein compartmentalization in the ICM/TE was also studied at E7 (Day 7 blastocyst), E8 (Day 8 blastocyst) and E9 (Day 9 blastocyst) as well. Encircled areas demarcate the ICM.

Article Snippet: NANOG was labelled with a rabbit polyclonal anti-NANOG antibody (Reprotech; 1∶500 in PBS–BSA for immunofluorescence and 1/1000 in PBS–T 4% non-fat dried milk for Western blot).

Techniques: Immunofluorescence, Western Blot

Journal: PLoS ONE

Article Title: Expression of Pluripotency Master Regulators during Two Key Developmental Transitions: EGA and Early Lineage Specification in the Bovine Embryo

doi: 10.1371/journal.pone.0034110

Figure Lengend Snippet: Expression patterns of OCT4 , SOX2 and NANOG were determined in bovine cloned morulae (Clone Morula A and Clone Morula B) and were compared with the levels of expression of these genes in controls (IVF Morulae). The results of qRT-PCR in all three types of morulae were normalised using an exogenous transcript luciferase. Values are means ± SEM of one equivalent embryo. The experiment was repeated three times and each repetition contained a triplicate of each sample.

Article Snippet: NANOG was labelled with a rabbit polyclonal anti-NANOG antibody (Reprotech; 1∶500 in PBS–BSA for immunofluorescence and 1/1000 in PBS–T 4% non-fat dried milk for Western blot).

Techniques: Expressing, Clone Assay, Quantitative RT-PCR, Luciferase