Fig. 5: Transcriptional activity of oyster cgPdx and cgNeuroD in cell culture and oyster hepatopancreas tissue.

a Transcriptional activity of oyster cgPdx on cgILP B1 element in COS7 cells demonstrating dose-dependent induction of luciferase reporter gene expression. Points show values of each individual assay. Violin and box plots are used to show the distribution of the data, with the mean at the right side. p values of comparisons (two-sided t-test) between adjacent groups are shown. b Transcriptional activity of cgNeuroD on cgILP B1 element in COS7 cells showing a similar but weaker dose-dependent induction effect. c Co-transfection of cgPdx and cgNeuroD gives higher transcriptional activity than the sum of the individual activities, suggesting functional synergy between cgPdx and cgNeuroD proteins. 150 ng of cgPdx and cgNeuroD plasmids was used for group (cgPdx + cgNeuroD). The pool of all the possible sums of individual relative luciferase values between cgNeuroD and cgPdx groups was used to estimate the 95% confidence interval (CI) of the activity sum. Significance of difference between group (cgPdx + cgNeuroD) and the sum of cgNeuroD and cgPdx groups was calculated by bootstrapping (n = 100,000). For experiments in a–c, n = 3 biologically independent cells by two technically independent measurements. Box represents a range from the first to the third quartile while whisker shows the minimum and maximum values. The line in each box shows the median. Relative luciferase activity in the cells transfected with blank pSI and the reporter plasmid is set at 1.00. p values were calculated by two tailed Student’s t test and Holm’s method to correct for multiple comparisons (p.adj). d–f ChIP-qPCR assays on predicted cgPdx binding sites upstream of three putative Pdx target genes in oyster (cgILP, cgPCSK1, and cgXBP1). Two anti-cgPdx polyclonal antibodies (E9112 and E9113) were used to assay two oyster hepatopancreas samples (Hep1 and Hep2). Source data are provided as a Source Data file.