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Commit 03971f17 authored by Benedikt Venn's avatar Benedikt Venn
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...@@ -24,6 +24,28 @@ Comparative analysis of both experiments were performed within this ARC. These i ...@@ -24,6 +24,28 @@ Comparative analysis of both experiments were performed within this ARC. These i
- visualization of all transcript signals and combined functional terms (Fig. 1) - visualization of all transcript signals and combined functional terms (Fig. 1)
<img src="https://https://git.nfdi4plants.org/lux/carbon-availability-transcriptomics-chlamy/-/raw/main/studies/HeatstressExperiment/resources/Figure01.png" alt="Signal" width="80%"/> <img src="https://git.nfdi4plants.org/lux/carbon-availability-transcriptomics-chlamy/-/raw/main/studies/HeatstressExperiment/resources/Figure01.png" alt="Signal" width="80%"/>
_Figure 1: Example of generated figures. **(A)** Exemplary visualization of the normalized counts of the HSP70C transcript. A clear separation of the different temperature kinetics is visible. While the initial level is comparable for all time courses after heat onset of 35°C transcript counts increase strongly while the 25°C signal seems constant. Transcript counts in both 40°C experiments decreased during the first 4 hours of treatment. After 8 hours of heat stress the behaviour of temperature-regulated signals change to medium specific effects. Cells living in low-acetate media show distinct reduction of HSP70C transcripts while TAP-samples settle approximately at prior-heat levels. **(B)** Heatmap representation of (A). **(C)** Transcript signals that belong to the functional term "intraflagellar transport.IFT particle protein.complex B" are visualized as z-scores. Since there was no measurement for TAP-25°C this panel remains empty (lower left). Because z-scores may distort signals that did not change at all, a ANOVA was performed to separate constant transcript from transcripts that showed differential expression within their time courses. Red shadings indicate a global change of the transcript counts. Blue/grey signals can be considered as constant and have less relevance for the shown kinetics. As seen most transcripts show no response at 25°C but show distinct patterns for 35°C and 40°C respectively. The response shape is solely dependent on the applied temperature and is not influenced by the media the cells are grown in. While for 35°C the transcripts show a strong decrease within the first two hours and a strong increase during the last period, for 40°C samples the transcripts remain constant for the first 2 hours and show no strong increase for the last time point._
References:
- Analysis:
- Benedikt Venn, Lukas Weil, Kevin Schneider, David Zimmer & Timo Mühlhaus. (2022). fslaborg/FSharp.Stats. Zenodo. https://doi.org/10.5281/zenodo.6337056
- Kevin Schneider, Lukas Weil, David Zimmer, Benedikt Venn & Timo Mühlhaus. (2022). CSBiology/BioFSharp. Zenodo. https://doi.org/10.5281/zenodo.6335372
- Love, M.I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15, 550 (2014). https://doi.org/10.1186/s13059-014-0550-8
- Visualization:
- Schneider K, Venn B and Mühlhaus T. Plotly.NET: A fully featured charting library for .NET programming languages [version 1; peer review: awaiting peer review]. F1000Research 2022, 11:1094 (https://doi.org/10.12688/f1000research.123971.1
- Data and annotation:
- Merchant, S. S., Prochnik, S. E., Vallon, O., Harris, E. H., Karpowicz, S. J., Witman, G. B., … Grossman, A. R. (2007). The Chlamydomonas Genome Reveals the Evolution of Key Animal and Plant Functions. Science, 318(5848), 245–250. https://doi.org/10.1126/science.1143609
- Zhang, N., Mattoon, E.M., McHargue, W. et al. Systems-wide analysis revealed shared and unique responses to moderate and acute high temperatures in the green alga Chlamydomonas reinhardtii. Commun Biol 5, 460 (2022). https://doi.org/10.1038/s42003-022-03359-z
_Figure 1: Example of generated figures. **(A)** Exemplary visualization of the normalized counts of the HSP70C transcript. A clear separation of the different temperature kinetics is visible. While the initial level is comparable for all time courses after heat onset of 35°C transcript counts increase strongly while the 25°C signal seems constant. Transcript counts in both 40°C experiments decreased during the first 4 hours of treatment. After 8 hours of heat stress the behaviour of temperature-regulated signals change to medium specific effects. Cells living in low-acetate media show distinct reduction of HSP70C transcripts while TAP-samples settle approximately at prior-heat levels. **(B)** Heatmap representation of (A). **(C)** Transcript signals that belong to the functional term "intraflagellar transport.IFT particle protein.complex B" are visualized as z-scores. Since there was no measurement for TAP-25°C this panel remains empty (lower left). Because z-scores may distort signals that did not change at all, a ANOVA was performed to separate constant transcript from transcripts that showed differential expression within their time courses. Red shadings indicate a global change of the transcript counts. Blue/grey signals can be considered as constant and have less relevance for the shown kinetics. As seen most transcripts show no response at 25°C but show distinct patterns for 35°C and 40°C respectively. The response shape is solely dependent on the applied temperature and is not influenced by the media the cells are grown in. While for 35°C the transcripts show a strong decrease within the first two hours and a strong increase during the last period, for 40°C samples the transcripts remain constant for the first 2 hours and show no strong increase for the last time point._
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## Normalization
Merged count tables from TAP and nopump samples were normalized together for further comparison/analysis.
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