Journal: ACS chemical neuroscience

Article Title: Passage Variation of PC12 Cells Results in Inconsistent Susceptibility to Externally Induced Apoptosis

doi: 10.1021/acschemneuro.6b00208

Figure Lengend Snippet: Optimization and validation of etoposide-induced cell death in PC12 cells (passage 9–13). 50 × 103 cells/well. (a) Approximately 50% cell death was observed after 48 h. Four replicates per experiment. Values are represented as mean ± SD. Multiple t test, Holm-Sidak method, alpha = 5.000%, ****p < 0.0001. (b) Fluorescence measured after 48 h. Flupirtine at 3 μM concentration rescued the cells from apoptosis. Experiment repeated thrice. Values are represented as mean ± SD. Unpaired two-tailed t test, 95% CI, ****p < 0.0001 in comparison to vehicle. ####p < 0.0001 in comparison to etoposide treatment.

Article Snippet: 5 Etoposide (Chem-Impex International, 28435) was stored as a working stock solution of 150 μ g/mL at −20 °C for up to 3 months.

Techniques: Fluorescence, Concentration Assay, Two Tailed Test, Comparison

Journal: ACS chemical neuroscience

Article Title: Passage Variation of PC12 Cells Results in Inconsistent Susceptibility to Externally Induced Apoptosis

doi: 10.1021/acschemneuro.6b00208

Figure Lengend Snippet: NGF differentiated PC12 cells show a statistically significant difference in sensitivity to apoptosis induced by etoposide between early and late passages at 72 h post insult. (a) Passage 10. Three replicates per experiment. Values are represented as mean ± SD. One-way ANOVA, Dunnett test, 95% CI. ****p < 0.0001. (b) Passage 17. Three replicates per experiment. Values are represented as mean ± SD. One-way ANOVA, Dunnett test, 95% CI. ****p < 0.0001. (c) Comparison of 72 h etoposide treatment in passage 10 and 17 PC12 cells differentiated using NGF. Unpaired two-tailed t test, 95% CI, ****p < 0.0001.

Article Snippet: 5 Etoposide (Chem-Impex International, 28435) was stored as a working stock solution of 150 μ g/mL at −20 °C for up to 3 months.

Techniques: Comparison, Two Tailed Test

Journal: The EMBO Journal

Article Title: Mitotic checkpoint gene expression is tuned by codon usage bias

doi: 10.15252/embj.2021107896

Figure Lengend Snippet: A Cellular protein noise (coefficient of variation, CV = std / mean) in live‐cell microscopy images of S. pombe ; n = 7 images (Nmt1‐GFP), 11 (Mad1‐GFP), 19 (Mad2‐GFP), 10 (Mad3‐GFP); single images had 16–79 GFP‐positive and 6–94 GFP‐negative (control) cells. Boxplots show median and interquartile range (IQR); whiskers extend to values no further than 1.5 times the IQR from the first and third quartile, respectively. Mad1, Mad2, and Mad3 all showed significantly lower noise than Nmt1 (Wilcoxon rank sum test; all P < 0.001). B Simulations of stochastic gene expression noise from selected mRNA/protein half‐life combinations assuming a constantly active promoter (see Methods). Synthesis rates were set to obtain a mean mRNA number of 4 per cell, and a mean protein number of 6,000 per cell. The x‐axis of each graph shows time, the y‐axis shows mRNA number per cell (blue) or protein number per cell (black). C Theoretical prediction for the coefficient of variation (CV = std/mean) of the protein number per cell, assuming different mRNA and protein half‐lives, using the same underlying model as in B. Synthesis rates were adjusted to maintain a mean mRNA number per cell of 3.5, and a mean protein number per cell of 6,000 (approx. 100 nM). D mRNA abundances by qPCR following metabolic labeling and removal of the labeled pool (two independent experiments). Lines are regression curves from generalized linear mixed model fits, excluding the measurements at t = 0 in order to accommodate for noninstantaneous labeling by 4tU. Act1 + and ecm33 + were used as long and short half‐life controls, respectively; qPCR was performed for the endogenous mRNAs. Half‐lives (95% confidence interval): mad1 + 5.6 min (4.3–8.4), mad2 + 7.7 min (6.2–10.4), mad3 + 5.2 min (4.3–6.9), act1 + 61.8 min (37.2–172.3), ecm33 + 5.0 min (4.5–5.7). E Protein abundances after translation shut‐off with cycloheximide (CHX); n = 3 experiments, error bars = std. Lines indicate fit to a one‐phase exponential decay. Cdc2 and Cdc13 were used as long and short half‐life controls, respectively. Immunoblots for the endogenous proteins (no tag). A representative experiment shown in Appendix Fig . Source data are available online for this figure.

Article Snippet: Cycloheximide (from Streptomyces griseus) , Chem Impex , Cat # 00083.

Techniques: Microscopy, Negative Control, Expressing, Labeling, Western Blot

Journal: The EMBO Journal

Article Title: Mitotic checkpoint gene expression is tuned by codon usage bias

doi: 10.15252/embj.2021107896

Figure Lengend Snippet:

Article Snippet: Cycloheximide (from Streptomyces griseus) , Chem Impex , Cat # 00083.

Techniques: Recombinant, In Vitro, Sequencing, Labeling, Protease Inhibitor, Isolation, Magnetic Beads, Bicinchoninic Acid Protein Assay, Software, Membrane