Additional file 2: Fig. S1. of Genome-wide miR-155 and miR-802 target gene identification in the hippocampus of Ts65Dn Down syndrome mouse model by miRNA sponges

a) Schematic representation of lentiviral vectors used to overexpress mmu-miR-155 and mmu-miR-802 (p-miR155 and p-miR802). b) Expression of mmu-miR155 and miR-802 in HeLa cells transfected with p-miR155 and p-miR802 (HeLa-miR-155-miR802). c) d2EGFP expression in HeLa and HeLa-miR-155-miR802 transduced with Lv-Control Lv-miR155T or Lv-miR155-802T. Top panel shows representative images of d2EGFP positive cells at 72-hours after transduction with 10 MOI of Lv-Control, LvmiR155Tor Lv-miR155-802T. Bottom panel shows the number of d2EGFP positive cells at different viraldoses. Image analysis was assessed using ImageJ software. Fig. S2. a) Principal component analysis of data obtained with the Agilent SurePrint G3 Mouse gene expression 8x60K Microarray (ID 028005). TheGEO accession number for our microarray data reported in this paper is GSE68074. Table of meandistances among the different genotypes and treatments. b) Prediction of miRNA targets based oncorrelation coefficients obtained from hippocampus treatment with sponge lentiviruses by different methods.Left panel shows the correlation prediction compared to the number of target sites predicted by TargetScan.Central panel shows the correlation prediction compared to the probability of conserved targeting(TargetScan PCT score). Right panel shows the correlation prediction compared to TargetScan contextscore. Fig. S3. a) Representation of Down’s syndrome gene expression dysregulation domanins (GEDDs)described by Letourneau and collaborators (black barplot overlay), and changes in expression in the miceorthologous genes observed in hippocampus upon the treatment with Lv-miR155-802T (grey barplotoverlay). Representation was performed using CIRCOS visualization package. b) Venn diagram of miRNApredicted targets on Lv-miR155-802T treated hippocampus based on the negative coefficients obtainedfrom miRComb analysis package for miR-155 (Corr_155 -) and miR-802 (Corr_802 -), genes that presenteda negative fold-change in between euploid and trisomic condition (TS/WT -) and genes that presented apositive fold-change in tisomic cells upon the XIST-induced HSA21 silencing (TS-XIST/TS +). (DOCX 3411 kb)