Adenoviral vector transduced in vitro differentiated C57bl6 BMDM cells

Total mRNA was extracted at 48 hour post transduction using Qiagen RNeasy Kits following the manufacture’s protocol.1 ug ofRNAwas used for cDNA library construction and bulk RNA sequencing were performed by Novogene (Sacramento CA). RNA-seq raw data were analysised using Partek Flow. Briefly, sequencing reads were aligned to the mouse or human genome using Spliced Transcripts Alignment to a Reference (STAR 2.5.3a). Aligned reads were then quantified to transcriptome (mm10; Ensembl Transcripts release 100), and normalized (Median Ratio normalization). Differentiated gene expression analysis (DESeq2) were performed to generate raw counts in vitro differentiated murine bone marrow derived macrophages on differentiation day 5 were replated, and transduced with mock (untreated), adenovirus carring GFP (empty vector) or HER2 targeting therapeutic payload (HER2.eSPR) on day 6. cells were collected on day 10 for RNA extraction.