Our finding that B16LS9 cells respond directly to entolimod treatment by NF-κB activation (Supplementary Figure S1) indicates that entolimod might be expected to have some antitumor effect through TLR5 stimulation on B16LS9 tumor cells independently on TLR5 expression in tumor microenvironment, as was previously demonstrated in several other TLR5-positive tumor models [18, 21, 22]. This evidence concerns the gene NFKB1 and neoplasm.