Extended Data Fig. 3: IFN-α therapy induces TLR7/8 expression on myeloid cells.
a, Differential expression of Tlr7 in myeloid cells identified by scRNA-Seq in skin-draining lymph nodes after IFN-α treatment (10.000 U, 4 h, s.c.) of mice. Gene expression data extracted from https://www.immunedictionary.org/app/home. b, Tlr7 expression in sorted myeloid cells analyzed by Microarray. Gene expression data extracted from: http://rstats.immgen.org/Skyline_microarray/skyline.html?datagroup=IFN. IFN-α treatment: (10.000 U, 2 h). c, DCs were generated from bone marrow that was supplemented with FLT3L and Tlr8 mRNA expression was analyzed using qRT–PCR. Cells were treated for 24 h with IFN-α (500 or 5000 U/mL), or with IMQ (2.5 µg/mL). In c, d n = 3 mice per group from 1 experiment. d, Tlr8 mRNA expression was assessed in murine bone-marrow-derived macrophages using qRT–PCR. Macrophages were treated as described in (c). e, Heatmap depicts TLR7 expression on myeloid cells within tumor-draining lymph nodes. Treatments are indicated in color-coded rectangles above the heatmap. The fold change of the geometric mean fluorescence intensity (gMFI) of TLR7 on indicated myeloid cells of treated mice compared to the control is shown. In e–g n = 3 mice per group from 1 experiment. f, Spleens were analyzed for TLR7 expression. Heatmap was plotted as described in (e). g, Small intestines were analyzed for TLR7 expression. Heatmap was plotted as described in (e). h, Tlr8 mRNA expression plotted against a human myeloid cell signature gene (Cd1c) in a subset of melanoma patients included in the TCGA database. The subset consisted of patients who received IFN-α prior to biopsy (n = 29) and a control group who did not receive IFN-α (n = 14). i, Correlation plot of Tlr8 to Cd68 in a subset of melanoma patients from the TCGA database, as described in (h). Bar graphs are plotted as mean ± SEM. Dots in c and d represent biological replicates. Correlation is shown in a xy-plot with a linear regression. Heatmaps are color-coded (blue = low, red = high value). P values were calculated using unpaired, two-tailed t-tests (e–g), one-way ANOVA with Tukey´s post-test (c, d) and two-tailed Pearson´s correlation test (h, i). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.
