Synthetic sickness of <i>swi/snf nhp6ΔΔ</i> and <i>rsc nhp6ΔΔ</i> triple mutants is suppressed by <i>2 µm-MRN1</i>.

<p>(A) and (B) Cells ten-fold serially diluted, spotted on SC plates and incubated for four days. Wild type: SG632; <i>swi2Δ</i>: SG418; <i>nhp6ΔΔ</i>: SG727; <i>swi2Δ nhp6ΔΔ</i>: SG759; wild type: SG358; <i>rsc8-ts21</i>: SG359; <i>nhp6ΔΔ</i>: SG394; <i>rsc8-ts21 nhp6ΔΔ</i>: SG658. Colony rows compared in the same panel derives from one plate. (C) Ability of <i>2 µm-MRN1</i> to suppress <i>rsc8-ts16 nhp6ΔΔ</i>. Cells streaked on SC-His plates and incubated for four days. Shown on the plates are two transformants containing <i>2 µm</i>-vector and four transformants containing <i>2 µm-MRN1</i>. <i>rsc8-ts16 nhp6ΔΔ</i>: SG657; <i>2 µm</i>-vector: pTK839; <i>2 µm-MRN1</i>: pTK1395. (D) Western blot analysis to visualize levels of endogenously expressed and <i>2 µm</i> expressed Mrn1-Myc. Rpb3-HA serves as a loading control. Untagged strain TG694 and tagged strain SG640 containing either pTK839 or pTK1423. Two µg of whole cell extract was separated on a SDS-PAGE and transferred to a mixed cellulose ester membrane and immunoblotted with anti-HA or anti-Myc antibody as indicated. (E) A schematic representation of the predicted domains and identified regions in Mrn1 (See text for details).</p

Plasmids Used in This Study.

<p>Plasmids Used in This Study.</p

Oligonucleotides Used in This Study.

<p>Oligonucleotides Used in This Study.</p

<i>rsc8-ts16 nhp6ΔΔ</i> cells accumulate unspliced transcripts.

<p>(A) Northern blot analysis was done with total RNA isolated from logarithmically SC-His growing cells at 25°C or after a two hour shift at 37°C. Total RNA was electrophoresed in a 0.25 M formaldehyde agarose gel, blotted and hybridized with specific ³²P-labeled probes. The probe was either intron-specific or 3′ exon-specific, respectively, for the <i>ECM33</i> RNA (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044373#pone.0044373.s001" target="_blank">Figure S1</a>). Ethidium bromide staining of the 18S and 25S rRNA is shown as a loading control. (B) Northern blot analysis was done with total RNA isolated from logarithmically SC-His growing cells at 30°C or after a two hour shift at 37°C. The probe was specific for both the <i>RPS11B</i> pre-mRNAand for the <i>RPS11B</i> mRNA (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044373#pone.0044373.s001" target="_blank">Figure S1</a>). Ethidium bromide staining of the 18S and 25S rRNA is shown as a loading control. (C), (D) and (E) Total RNA isolated from logarithmically SC-His growing cells at 25°C or after a two-hour shift at 37°C amplified by RT-qPCR with <i>ECM33-</i>, <i>ACT1-</i>, <i>ASC1-</i>, <i>RPS11B-</i> or <i>RDN25-</i>specific primers. (C) The ratio intron-3′exon junction RT-PCR-amplificate/3′exon RT-PCR-amplificate. (D) The ratio 3′exon RT-PCR-amplificate/<i>RDN25</i> RT-PCR-amplificate. (E) The ratio intron-3′exon junction RT-PCR-amplificate/<i>RDN25</i> PCR-amplificate. The ratio in wild type cells at 25°C was arbitrarily set to 1. Wild type: SG632; <i>rsc8-ts16 nhp6ΔΔ</i>: SG657; <i>2 µ</i>-vector: pTK839; <i>2 µ-MRN1</i>: pTK1423.</p

Knowledge deficit, attitude and behavior scales association to objective measures of sun exposure and sunburn in a Danish population based sample

<div><p>The objective of this study was to develop new scales measuring knowledge and attitude about UVR and sun related behavior, and to examine their association to sun related behavior objectively measured by personal dosimetry. During May-August 2013, 664 Danes wore a personal electronic UV-dosimeter for one week that measured their UVR exposure. Afterwards, they answered a questionnaire on sun-related items. We applied descriptive analysis, linear and logistic regression analysis to evaluate the associations between the questionnaire scales and objective UVR measures. Perceiving protection as routine and important were positively correlated with protective behavior. <i>Knowledge deficit of UV and risk of melanoma</i>, <i>perceived benefits</i> and <i>importance of protection behavior</i> was also correlated with use of protection. ‘<i>Knowledge deficit of UV and risk of melanoma</i> and <i>Perceived barrier towards sun avoidance between 12 and 15’</i> were both associated with increased risk of sunburn. <i>Attitude towards tan</i> was associated to both outdoor time and exposure as well as use of protection, but not to sunburn. The results regarding <i>Knowledge deficit of UV and risk of melanoma</i> associated to UVR exposure and <i>Perceived barrier towards sun avoidance between 12 and 15</i> emphasize the importance of awareness of melanoma risk and the priority of the skin cancer prevention advice. Shifting activities to outside the suns peak-hours could be an approach for structural and campaign preventive measures. Knowledge of items predicting exposure to UVR, use of protection and sunburn are important for planning of preventive interventions and melanoma research.</p></div

U4/U6 dimer, free U4 and total U6 snRNA levels in <i>rsc8-ts16 nhp6ΔΔ</i> cells.

<p>Total RNA prepared from logarithmically SC-His growing cells at 25°C or after a two-hour shift at 37°C. (A) RNA was fractionated on a non-denaturing 6% polyacrylamide gel, blotted and hybridized with a U4 specific probe. After analysis the membrane was stripped and re-probed with a U1 specific probe. (B) RNA was fractionated on a denaturing 6% polyacrylamide gel, blotted and hybridized with a U6 specific probe. After analysis the membrane was stripped and re-probed with a U1 specific probe. (C) Quantification of U4/U6 dimer, free U4 and total U6 snRNA amounts relative to U1 snRNA based on quantification of Storm Images from at least five individual experiments. Wild type: SG632; <i>rsc8-ts16 nhp6ΔΔ</i>: SG657.</p

Mrn1-GFP accumulates in the nucleus in a <i>mex67-5</i> mutant at 37°C.

<p><i>MRN1-GFP</i> and <i>mex67-5 MRN1-GFP</i> cells were harvested after growth in SC medium at 25°C or after 30 min incubation at 37°C. For each genotype and growth condition, 100–200 cells were inspected. <i>Error bars</i> indicate 95% confidence intervals. SG1008: <i>mex67-5 MRN1-GFP ADH1p-NLS-yEmRFP::URA3</i> and SG1010: <i>MRN1-GFP ADH1p-NLS-yEmRFP::URA3</i>.</p

Yeast Strains Used in This Study.

<p><i>MAT</i>¹: The mating type has not been determined.</p

Genetic interactions linking <i>MRN1</i> and chromatin mutants to pre-mRNA splicing.

<p>(A) <i>mrn1Δ</i> is synthetic sick with <i>swi2Δ</i>. Cells ten-fold serially diluted, spotted on SC plates or SC plates containing 3% formamide and incubated for four days at 30°. <i>swi2Δ</i>: SG418; <i>mrn1Δ</i>: SG520; wild type: SG632 and <i>mrn1Δ swi2Δ</i>: SG766. (B) <i>mrn1Δ</i> is synthetic sick with <i>nhp6ΔΔ</i>. Cells ten-fold serially diluted, spotted on SC plates and incubated for four days at the indicated temperatures. <i>mrn1Δ</i>: SG520; wild type: SG632; <i>nhp6ΔΔ</i>: SG727 and <i>mrn1Δ nhp6ΔΔ</i>: SG762. (C) <i>mrn1Δ snt309Δ</i> cells are synthetic sick. Cells ten-fold serially diluted, spotted on SC plates and grown at the indicated temperatures for four days. <i>mrn1Δ</i>: SG912; wild type: SG632; <i>snt309Δ</i>: SG648; <i>mrn1Δ snt309Δ</i>: SG920. (D) The temperature sensitivity of <i>snt309Δ</i> is suppressed by <i>2 µm-MRN1</i>. Cells ten-fold serially diluted, spotted on SC-His plates and incubated for four days at the indicated temperatures. Wild type: SG632; <i>snt309Δ</i>: SG648; <i>2 µm</i>-vector: pTK839; <i>2 µm-MRN1</i>: pTK1395. (E) The temperature sensitivity of <i>prp22</i> is suppressed by <i>2 µm-MRN1</i>. Cells ten-fold serially diluted, spotted on SC-His plates and incubated for four days at the indicated temperatures. Wild type: SG682; <i>prp22</i>: SG840; <i>2 µm</i>-vector: pTK839; <i>2 µm-MRN1</i>: pTK1423. (F) The temperature sensitivity of <i>prp4-1</i> is suppressed by <i>2 µm-MRN1</i>. Cells ten-fold serially diluted, spotted on SC-Ura plates and incubated for four days at the indicated temperatures. <i>prp4-1</i>: SG845; <i>2 µm</i>-vector: pTK51; <i>2 µm-MRN1</i>: pTK1386. (G) <i>snf5Δ</i> and <i>rsc2Δ</i> genetically interacts with <i>snt309Δ</i>. Cells ten-fold serially diluted, spotted on SC plates and incubated for four days at the indicated temperatures. <i>rsc2Δ</i>: SG417; <i>snf5Δ</i>: SG420; wild type: SG632; snt309Δ: SG729; rsc2Δ <i>snt309Δ</i>: SG773 and <i>snf5Δ snt309Δ</i>: SG774. (H) <i>snt309Δ</i> is synthetic lethal with <i>nhp6ΔΔ</i>. Cells ten-fold serially diluted, spotted on SC-Ura plates or 5-FOA plates and incubated for four days at 30°C. Wild type: SG865; <i>nhp6ΔΔ</i>: SG867; <i>snt309Δ</i>: SG868; <i>snt309Δ nhp6ΔΔ</i>: SG869; <i>2 µm</i>-<i>NHP6B-URA3</i>: pTK1382. Colony rows compared in the same panel derives from one plate.</p

Distribution of demographic characteristics and scale scores in a cross-sectional sample of 664 Danes.

<p>Distribution of demographic characteristics and scale scores in a cross-sectional sample of 664 Danes.</p