Journal: Journal of Aging Research

Article Title: Self-Renewal Signalling in Presenescent Tetraploid IMR90 Cells

doi: 10.4061/2011/103253

Figure Lengend Snippet: Antibodies: source and use.

Article Snippet: Rabbit polyclonal anti-hOCT4 (ab19857, Abcam) 1 : 200 , Goat anti-rabbit-IgG-Alexa Fluor 594 (A31631, Invitrogen).

Techniques:

Journal: Traffic (Copenhagen, Denmark)

Article Title: The Ubp3/Bre5 deubiquitylation complex modulates COPII vesicle formation

doi: 10.1111/tra.12766

Figure Lengend Snippet: The absence of the Ubp3/Bre5 complex reverts Kar2 secretion as a result of a reduction in ER‐to‐Golgi transport. A, Extracellular Kar2 was detected by colony overlay immunoblot in the indicated yeast strains. Serial dilutions of cells at stationary phase were spotted onto YPD plates. After ~5 hours growth, colonies were overlaid with nitrocellulose and secreted Kar2 detected with Kar2‐specific antibodies. B, Exponentially growing wild‐type and mutant cells carrying a UPRE ‐ LacZ reporter gene were lysed, and ß‐galactosidase activity measured using ONPG as a substrate. Error bars depict mean and SD of at least three independent experiments. Statistical test was a one‐way ANOVA with Dunnett's multiple comparisons test. C, The indicated strains were grown to exponential phase, serially diluted and spotted on media containing 5‐fluoroorotic acid (5‐FOA) to counter‐select for the wild‐type p SEC13 :: URA3 plasmid. D, Wild type, emp24Δ, emp24Δ ubp3Δ and emp24Δ bre5Δ cells were subjected to pulse‐chase analysis with [ 35 S] methionine. Gas1 was immunoprecipitated from cell lysates at the indicated times. Immunoprecipitated proteins were detected by SDS‐PAGE/autoradiography (left panel). Maturation of Gas1 was quantified by phosphorimage analysis (right panel) (E) Heat‐map showing the Kar2 reversion index of ubp3Δ and bre5Δ in different Kar2 secretor strains involved in a wide variety of biological processes

Article Snippet: Serial dilutions (1:10) were made from the saturated cultures and spotted onto solid media without (control) or with 0.1% 5‐Fluoroorotic acid (5‐FOA, BioVision) to counter‐select for the corresponding URA3 plasmid (p SEC13‐URA3 , p SEC23‐URA3 or p SEC24‐URA3 ).

Techniques: Western Blot, Mutagenesis, Activity Assay, Plasmid Preparation, Pulse Chase, Immunoprecipitation, SDS Page, Autoradiography

Journal: Traffic (Copenhagen, Denmark)

Article Title: The Ubp3/Bre5 deubiquitylation complex modulates COPII vesicle formation

doi: 10.1111/tra.12766

Figure Lengend Snippet: Genetic interactions between COPII proteins and Ubp3/Bre5. A, Plasmids encoding SEC23 mutants ‐ sec23‐1 and sec23‐Y678,684A ‐ were introduced into sec23Δ and sec23Δ ubp3Δ as indicated and grown on 5‐FOA at 25°C. B, Plasmids encoding SEC24 mutants ‐ sec24‐A, sec24‐C, sec24‐D and sec24‐m11 ‐ were introduced into sec24Δ and sec24Δ ubp3Δ as indicated and grown on 5‐FOA at 30°C

Article Snippet: Serial dilutions (1:10) were made from the saturated cultures and spotted onto solid media without (control) or with 0.1% 5‐Fluoroorotic acid (5‐FOA, BioVision) to counter‐select for the corresponding URA3 plasmid (p SEC13‐URA3 , p SEC23‐URA3 or p SEC24‐URA3 ).

Techniques: