Analysis from the results of a label free mass spectrometry experiment on paraffin embedded samples stratified as follows:
The coverage analysis result was conducted here on both data sets, and the summary statistics of the coverage score for each sample is shown on the tables below, and based on that and the plots we can see a similar behaviour regarding the coverage.
| rbc1 | rbc2 | rbc3 | rbc4 | mix1 | mix2 | mix3 | fib1 | fib2 | fib3 | fib4 | fib5 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Min. | 0.13000 | 0.25000 | 0.24000 | 0.05000 | 0.04000 | 0.30000 | 0.04000 | 0.45000 | 0.6500 | 0.26000 | 0.15000 | 0.19000 |
| 1st Qu. | 4.03500 | 5.13000 | 4.57000 | 4.47000 | 4.54000 | 4.11250 | 4.28000 | 5.03000 | 5.3725 | 3.28250 | 4.28000 | 4.79500 |
| Median | 8.95000 | 11.00000 | 9.59000 | 10.29000 | 9.75000 | 8.73000 | 9.72000 | 10.31000 | 7.1800 | 7.31500 | 9.78000 | 9.57000 |
| Mean | 14.02422 | 14.88716 | 14.31953 | 14.42899 | 14.78163 | 13.55094 | 14.25736 | 15.23341 | 15.4750 | 11.51224 | 14.81137 | 15.98963 |
| 3rd Qu. | 19.26000 | 20.05000 | 18.89000 | 19.16000 | 20.30000 | 17.94250 | 19.76000 | 21.08750 | 13.2950 | 14.51500 | 19.21000 | 22.23500 |
| Max. | 97.28000 | 97.28000 | 97.28000 | 97.28000 | 100.00000 | 97.28000 | 97.28000 | 66.90000 | 87.7600 | 68.47000 | 95.92000 | 63.27000 |
The Venn Diagram generated using the second data set shows that 101 proteins are overlapping between the RBC and FIB Groups - and this intersection is used for comparison between groups, 60 proteins are unique to the RBC samples, and 115 proteins are unique to the FIB samples.
Using the correlation matrix above to extract information on the abundancy profile of the top 50 highly correlated proteins with a threshold of +/- 0.80, the network analysis was carried out and resulted in the graph below. The node size is proportional with the degree (how connected the protein is), the red edges represent positive correlation and blue edges represent negative correlation. The optimal community structure was calculated for the graph, in using the maximal modularity score.This analysis resulted in the six communities represented as the node colours and shown in the table below.
The same method for constructing the co-expression network used on the RBC samples was applied to the FIB dataset here, in this case using the 115 proteins unique to FIB samples from the second batch. The optimal community analysis resulted in three communities listed in the table below.
Here I used a linear model approach to assess differential abundance/expression between the two groups - this analysis resulted in 40 differentially abundant proteins. The tables below show the metrics of the top-ranked proteins from the linear model fit. The data was pre-processed with log-transformation and quantile normalisation to ensure that the expression distributions of each sample are similar across the entire experiment and not skewed.