Additional file 1 of 7aaRGD - a novel SPP1/integrin signaling-blocking peptide reverses immunosuppression and improves anti-PD-1 immunotherapy outcomes in experimental gliomas

Additional file 1: Fig. S1 shows gene expression data of RGD motif-binding integrins in human glioma samples from public repositories and its correlation with patient survival. Fig. S2 presents the effect of the 7aaRGD peptide on gene expression in microglia-glioma co-cultures, shows representative pictures from Matrigel assay with two additional cell lines and presents the effect of the 7aaRGD peptide on the viability and proliferation of glioma and microglia cells. Fig. S3 describes stability studies and biodistribution of the 7aaRGD peptide delivered in osmotic pumps. Fig. S4 presents the profiles of myeloid cell in the brain of 7aaRGD-treated GL261 glioma-bearing mice analysed by flow cytometry and inferred from the deconvolution of bulk RNA-seq data of sorted tumor-infiltrating CD11b+ cells. Fig. S5 presents representative images from Gal3/ Tmem119 and Arg1 doublestaings. Fig. S6 shows extended data on characterization of the immune microenvironment of GL261 gliomas upon 7aaRGD or/and anti-PD-1 treatment. Fig. S7 presents the data on immune cells phenotyping in the TME and spleens at 21 DPI in glioma-bearing mice. Fig. S8 presents the profiles of intratumoral myeloid cells in different treatment groups inferred from the deconvolution of bulk RNA-seq data. Fig. S9 shows key parameters of the response to 7aaRGD-treatment in U87-MG-RFP + glioma-bearing mice. Fig. S10 and Fig. S11 present the gating strategy used for FACS-sorting and flow cytometry analysis, respectively. Table S1 lists all antibodies used for immunohistochemistry and flow cytometry stainings