Journal: Journal of cell science

Article Title: Error-prone meiotic division and subfertility in mice with oocyte-conditional knockdown of pericentrin.

doi: 10.1242/jcs.196188

Figure Lengend Snippet: Fig. 1. Oocyte-conditional knockdown of Pcnt disrupts aMTOC-associated protein localization and lowers female fertility. (A) Mean±s.e.m. number of viable pups per litter born to WT (n=3) and Tg (n=3) female mice mated to WT males during a 6-month period. (B) Representative images of prophase-I (GV-stage) and ovulated oocytes (MII) collected from WT and Tg females. DAPI-labeled DNA is shown in gray and Pcnt in red. Insets show 6× (a) and 3× (b,c) magnifications. Arrowheads denote misaligned chromosomes. G.Cell, granulosa cells. (C) Percentage (mean±s.e.m.) of total WT (n=79) and Tg (n=61) ovulated MII oocytes that show positive immunofluorescence Pcnt labeling. (D) Fluorescence intensity of Pcnt at aMTOCs in ovulated MII WT (n=20) and Tg (n=21) oocytes. (E) Relative transcript levels for Pcnt (Pcnt), γ-tubulin (Tubg1) and the γ-tubulin adaptor protein neural precursor cell expressed, developmentally down-regulated 1 (Nedd1) in ovulated oocytes (n=100) from control (WT) and transgenic (Tg) females. (F) Representative images of WT and Tg (Pcnt-depleted) oocytes, labeled with anti-Nedd1 (green) or anti-Cep215 (green). DAPI-labeled DNA is shown in gray. Arrowhead denotes misaligned chromosome. Scale bar of 10 μm. (G) Representative images of prophase-1 arrested WT and Tg oocytes, labeled with anti-γ-tubulin (green). The arrowhead denotes an aMTOC. Insets show a 6× magnification. (H) Percentage (mean±s.e.m.) of total WT (n=44) and Tg (n=42) prophase-I arrested oocytes that label positively for γ-tubulin at aMTOCs. (I) Representative images of ovulated MII WT and Tg oocytes, labeled with anti-γ-tubulin (green). Insets show a 2× magnification. Percentage (mean±s.e.m.) of total WT (n=90) and Tg (n=98) MII oocytes showing positive labeling for (I) γ-tubulin at the spindle poles and cytoplasmic aMTOC foci, or (J) γ-tubulin along the spindle microtubules (MTs). *P<0.05; ***P<0.001 relative to WT. Scale bars: 10 μm.

Article Snippet: Antibodies were used at a 1:1000 dilution to detect Pcnt (BD Biosciences and Covance, cat# PRB-432C), γ-tubulin (Sigma, Jo u rn al o f Ce ll Sc ie n ce cat# T3559), Nedd1 (Novus Biologicals, cat# H00121441-M05) and Cep215 (EMD, Millipore, cat# ABE236) as well as spindle MTs with acetylated α-tubulin (Sigma, cat# T6793, clone 6-11B-1).

Techniques: Knockdown, Labeling, Immunofluorescence, Fluorescence, Control, Transgenic Assay

Journal: eLife

Article Title: Self-assembly of pericentriolar material in interphase cells lacking centrioles

doi: 10.7554/eLife.77892

Figure Lengend Snippet:

Article Snippet: Antibody , anti-NEDD1 (rabbit polyclonal) , Rockland , Rockland Cat# 109–401 C38S, RRID: AB_10893219 , (1:1000) for WB.

Techniques: Sequencing, Luciferase, Imaging, Software, Recombinant, Plasmid Preparation, Knock-Out, Mutagenesis

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Involvement of centrosome amplification in radiation-induced mitotic catastrophe.

doi: 10.4161/cc.6.3.3834

Figure Lengend Snippet: Fig. 1. GFP-centrin-1 as a marker of IR- induced centrosome amplification. (A) Human U2OS cells expressing GFP-centrin-1 (green) and H2B-RFP (red) were fixed in methanol and stained with the following antibodies to centrosomal or centrosome-associated proteins, Aurora A, γ-tubulin, CEP170, Ninein and Nedd1 (blue, Alexa 350). Scale bar, 10 mm. U2OS cells as described in (A), before (B) and after (C) treatment with 10 Gy IR.

Article Snippet: Cells were fixed in either 4% PFA or methanol as described in ref. 22 and stained using the following primary antibodies: γ-tubulin (T3559, Sigma), ninein (AM), CEP170 (GG), Nedd1 (AM), Aurora A (35C1, Abcam), a-tubulin (B512, Sigma), TPX-2 (BL1679, Bethyl Labs), BubR1 (8G1, AbCam) and Mad2 (Abcam).

Techniques: Marker, Amplification, Expressing, Staining