Journal: PLoS Genetics

Article Title: The cAMP signaling pathway regulates Epe1 protein levels and heterochromatin assembly

doi: 10.1371/journal.pgen.1010049

Figure Lengend Snippet: (A) RT-qPCR analysis of epe1 + transcript, normalized to act1 + . Error bars represent SD, n = 2. (B, E) Western blot analysis to measure the levels of 3HA-tagged Epe1 under the control of nmt41 promoter and levels of tubulin. (C) Left, western blot analysis to measure the levels of 3HA-tagged Epe1 under the control of nmt41 promoter and tubulin. Samples are taken at indicated time points (minutes) after the addition of cycloheximide. Different exposures were used for wild type and git3Δ cells to make the signal at time 0 comparable. Right, quantification of Epe1 proteins levels. Error bars represent SD, n = 2. (D) Top, sucrose density gradients of ribosomes in each strain with continuous monitoring of absorbance at 260 nm. Lighter fractions are on the left. Bottom, qRT-PCR analysis of epe1 + , act1 + , and nmt1 + transcripts from each fraction. Relative amounts of transcripts were calculated using the delta Ct method. The distribution is shown as a percentage of the total. Error bars represent the SD of two technical replicates.

Article Snippet: Quantitative real-time PCR (qPCR) was performed with Luna Universal qPCR Master Mix (NEB, M3003X) in a StepOne Plus Real-Time PCR System (Applied Biosystems).

Techniques: Quantitative RT-PCR, Western Blot

Journal: PLoS Genetics

Article Title: The cAMP signaling pathway regulates Epe1 protein levels and heterochromatin assembly

doi: 10.1371/journal.pgen.1010049

Figure Lengend Snippet: (A, C) Western blot analysis to measure the levels of endogenous 3HA-tagged Epe1 and tubulin. (B) RT-qPCR analysis of epe1 + transcript levels, normalized to act1 + . Error bars represent SD. n = 2. (C) Sucrose density gradients of ribosomes in each strain with continuous monitoring of absorbance at 260 nm. Lighter fractions are on the left. (E) qRT-PCR analysis of epe1 + transcripts from each fraction. Relative amounts of transcripts were calculated using the delta Ct method. The distribution is shown as a percentage of the total. Error bars represent the SD of two technical replicates.

Article Snippet: Quantitative real-time PCR (qPCR) was performed with Luna Universal qPCR Master Mix (NEB, M3003X) in a StepOne Plus Real-Time PCR System (Applied Biosystems).

Techniques: Western Blot, Quantitative RT-PCR