Fig. 3: Transcriptomic and Protein Expression Analysis of ICAM-1 Knockout (KO) vs Wild Type (WT) Cells.

H9 ICAM-1 KO and WT isogenic pluripotent stem cells (PSCs) were stimulated with TNFα (10 ng/ml) and IFNγ (50 ng/ml) for 48 h, then assessed for (A) MHC class I expression and (B) surface ICAM-1 expression at baseline and following stimulation. Data representative of 3 independent experiments. MFI = Median Fluorescence Intensity. Analysis was performed via FlowJo v10 software. For A and B, the red and the blue lines indicate the MFI of the unstimulated and the stimulated PSCs, respectively. C Western blot was performed on the protein extracted from unstimulated H9 WT PSCs and from H9 WT and ICAM-1 KO PSCs stimulated with TNFα (10 ng/ml) and IFNγ (50 ng/ml) for 44 h. Each sample consisted of pooled biological replicates (BR) to achieve sufficient protein concentration. The stimulated H9 WT protein sample consisted of n = 5 BR, unstimulated H9 WT consisted of n = 2 BR, and H9 ICAM-1 KO consisted of n = 3 BR. L denotes ladder placement. D Bulk RNA sequencing was performed on PED05 ALG ICAM-1 KO vs. WT induced PSCs (iPSCs); PED05 ALG WT iPSC-derived cardiomyocytes (CMs) shown as a differentiated cell type reference control. Log2 RSEM (v1.2.3.1) counts for 21,055 transcripts from four BR of PED05 ICAM-1 KO iPSCs, four BR of PED05 WT iPSCs, and four BR of PED05 WT iPSC-derived CMs. Clustering was performed and the graphic was made by running pheatmap in R Studio (v4.4.1) using Euclidean distance and ward.D hierarchical clustering metrics. E Log2 RSEM counts for 60 gene reference markers classifying PSC identity. Other abbreviations – ALG Akaluc + GFP (green fluorescent protein) reporter construct. Source data are provided as a Source Data file.