Fig. 1
From: Alternative splicing is required for stage differentiation in malaria parasites
Flowcharts depicting transcriptional changes between stages and qRT-PCR verification. a Transcripts are upregulated in both female and male gametocytes or downregulated in both sexes; this comprises 13.1% of genes with detected changes in expression. The remaining genes have changes in their gene expression unique to female and/or male gametocytes. Arrow widths are proportional to the associated percentages, which are derived from the number of genes changing between those stages. (N.B., these do not total 100%, as genes upregulated in females and downregulated in males, or vice versa, are considered “unique” to both sexes). b When looking at the changes in alternative splicing, only 11.4% of genes with detected changes in alternative splicing are common to both gametocyte sexes. Two examples of female- and male-specific changes in alternative splicing are depicted. Gene models are to scale, reading sense left-to-right, with the middle model representing the primary isoform. Isoforms linked to specific stages are relatively enriched in that stage, with alternatively spliced regions highlighted. Black rectangles indicate protein-coding regions, gray boxes indicate untranslated regions, and lines indicate introns. c RNA-seq analyses revealed changes in alternative splicing between female and asexual parasites in the following genes, which were verified by qRT-PCR. The first four (red) genes were predicted to have a greater proportion of transcripts that retained the highlighted sub-exonic bin in females compared to asexuals (as indicated by ΔΔCt greater than zero), the next (blue) gene was predicted to have fewer transcripts in females with the highlighted sub-exonic bin (ΔΔCt less than zero), and the gray genes were predicted to have no changes in splicing (no difference in ΔΔCt). mRNA isoforms to scale are displayed on the left, reading sense left-to-right; sub-exonic bins with a predicted change are highlighted in red or blue. We observed statistical significance in the first five genes, as expected (p<0.05, detailed analysis in Additional file 3: Table S3)