Genes related to the TNF signalling pathway show switching between isoform expression with potential functional implications. (A) Based on the analysis of switching behaviour between isoforms (Figure 4A), we further investigated the potential functionality of the isoforms. The two isoforms of TNFRSF12A have different predicted functionality. ENST00000326577.8 has predicted coding functionality and a stn_TNFRSF12A domain, whereas the ENST00000341627.5 is predicted to be non-coding. (B) The CD44 gene with its multiple isoforms illustrates the importance of alternative splicing: the total gene expression is almost unchanged between the time points 30 h and 39 h, however the individual isoforms show a larger deviation in expression. Most notably, the ENST00000528086.5 isoform, predicted to be coding, is almost absent at 39 h. (C) In the L-1236 cell line we detected changes in isoform balance in the NFKB2 gene (Figure 4C). The total expression of the gene shows only a minor change between the time points, however the isoform ENST00000467116.5 lacking the important death-domain is upregulated at 30 h as compared to 24 h (STP2).