Figure 4. PEDD-nelitolimod in combination with Sys or SQ α-PD-1 promoted transcriptomic changes consistent with enhancement of antitumor immunity in the liver tumor microenvironment. Total RNA was isolated from the CD45⁺ cells of liver metastases, and nCounter analysis was performed using myeloid and innate panel. (A) Heat map of genes that were significantly upregulated/downregulated followed by α-PD-1 Ctrl, nelitolimod, Sys and SQ treatment compared with Veh control were plotted. (B) The Venn diagram compared genes significantly modulated by PEDD of nelitolimod/SQ/Sys compared with Veh. (C) i–ii Expression of Ifn-γ and granzyme were quantified by qRT PCR. (D) Pathways that were altered by nelitolimod, α-PD-1 monotherapy or combination therapy compared with Veh, evaluated by using nSolver advanced analysis module and plotted, respectively. Data presented as mean±SEM and n are mentioned in the individual graph. One-way analysis of variance was performed to determine statistical differences among multiple groups. qRT, quantitative reverse transcription; IFN, interferon; ECM, extracellular matrix; PEDD, pressure-enabled drug delivery; PD-1, programmed cell death-1; SQ, subcutaneous; Sys, systemic; Veh, vehicle.