Fig. 3

pcsk9 is up-regulated on a pneumococcal infection and associated with the expression of acute-phase reactants. A, The relative expression of pcsk9 was determined in the liver of PBS injected (n = 5; all males) and S pneumoniae infected (635 000 CFU; SD 276 000 CFU, n = 5; all males) WT AB zebrafish at 7 dpi using qPCR. Gene expression levels were normalized to eef1a1l1 expression and target genes were run once as technical duplicates. A 2-tailed Mann-Whitney test was used for statistics. B-C, RNA was isolated from the liver of unchallenged (n = 2 in both groups; 2 females/group), PBS injected (n = 3 in both groups; 3 females/group) and S pneumoniae infected (3 370 000 CFU; SD 840 000 CFU, n = 3 in both groups; 3 females/group) adult pcsk9^{tpu-13/tpu-13} and WT (pcsk9^tpu-13) zebrafish at 1 dpi and the transcriptome was analysed using RNA sequencing. B, Venn diagram for the differentially expressed genes within treatment groups. C, The top 25 up- and down-regulated genes in S pneumoniae infected pcsk9^{tpu-13/tpu-13} zebrafish in comparison with the WT controls are depicted using a heat-map. Statistics were done using DESeq2 and adjusted using the Benjamini-Hochberg (BH) method. D, The relative expression of selected genes up- or down-regulated in the RNA sequencing data (ldlrap1a, hamp and socs3a) was quantified using qPCR. Gene expression levels were normalized to eef1a1l1 expression and target genes were run once as technical duplicates