Fig. 3: Elevated UPP1 expression in tumor cells promotes immunosuppression through a TGF-β1-dependent manner. | Nature Communications

Fig. 3: Elevated UPP1 expression in tumor cells promotes immunosuppression through a TGF-β1-dependent manner.

From: UPP1 promotes lung adenocarcinoma progression through the induction of an immunosuppressive microenvironment

Fig. 3

a 80-cytokine array analysis between UPP1-overexpressing HCC827 tumor cells (UPP1-OE) and control cells (UPP1-OENC) (n = 3). b Top six upregulated cytokines in HCC827 UPP1-OE tumor cells (n = 3). c ELISA comparison of TGF-β1 expression in culture supernatants of HCC827 UPP1-OE and UPP1-OENC tumor cells (n = 4). d Co-culture workflow, created with BioRender.com. αTGF-β1, TGF-β1 neutralizing antibodies; IgG, isotype control IgG. e Flow cytometry analysis of CD25 + FOXP3+ Tregs proportion (n = 4). f Flow cytometry analysis of LAG3 + PDCD1 + CD8 + T cells proportion (n = 4). g Representative immunofluorescence (IF) staining of SPP1 in THP-1-derived macrophages (n = 4). MFIs represent mean fluorescence intensities. Scale bars = 20 μm, h Western blot analysis of SPP1 expression in THP-1-derived macrophages after co-culture (n = 3). i RT-qPCR measurement of M2 macrophage markers in THP-1-derived macrophages post co-culture (n = 3). j scRNA-seq analysis of fibroblasts after co-culturing with HCC827 UPP1-OE tumor cells/UPP1-OENC tumor cells. UPP1-OE tumor cells = 7707; UPP1-OENC tumor cells = 8200. k Comparison of CAFs scores in fibroblasts co-cultured with either UPP1-OE tumor cells or UPP1-OENC tumor cells. l Dot plot showing the mean expression of CAFs marker genes. m The UMAP plot displaying the distribution of the MMP11+ CAFs scores. Fibroblasts with an MMP11+ CAFs score higher than the median value were categorized as MMP11+ CAFs-like fibroblasts, while the others were defined as MMP11- fibroblasts (Fbs). n Comparison of the proportion of MMP11+ CAFs-like fibroblasts between the UPP1-OE tumor cells group and the UPP1-OENC tumor cells group. o Comparison of the CAFs-related biological processes between the UPP1-OE tumor cells group and the UPP1-OENC tumor cells group. p Western blot analysis of CAFs markers after the treatment of TGF-β1 neutralizing antibodies (n = 3). n denotes biologically independent samples. Data were represented as mean ± SD in b, c, e–g. Data were represented as mean ± SEM in i. Two-tailed student’s t-test was used in b, c; one-way ANOVA with multiple comparisons in e–g, i; two-tailed Wilcoxon rank-sum test in k and o. Two-tailed Fisher’s exact test in n. Source data are provided as a Source Data file.

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