Fig 2: S100A8 promotes BNIP3, BECN1 and PI3KC3 expression. (A) WB for BNIP3 in the endoplasmic reticulum of BCL cells following S100A8 interference. (B) WB for BNIP3 in the mitochondria of BCL cells following S100A8 interference. Co-immunoprecipitation assay for (C) BECN1-PI3KC3 and (D) BECN1-BCL2 complexes in Jeko-1 cells following S100A8 interference. BNIP3, Bcl-2/adenovirus E1B 19-kDa protein-interacting protein 3; BECN1, Beclin-1; PI3KC3, class III phosphatidylinositol-3-kinase; WB, western blotting; BCL, B-cell lymphoma.

Fig 3: S100A8 acts downstream of Sec23a to attenuate lung metastatic colonization of melanoma cells.a, b Representative images and quantification of transwell migration and invasion assay of tumor cells with S100A8 interference. Scale bars, 60 µm. *P < 0.05, **P < 0.01. c, d Representative images and quantification of colony formation assay of tumor cells with S100A8 interference. **P < 0.01. e, f Representative images and quantification of transwell migration and invasion assay of tumor cells treated with anti-S100A8/A9 antibody. Scale bars, 60 µm. **P < 0.01, ***P < 0.001. g, h Representative images and quantification of colony formation assay of tumor cells treated with anti-S100A8/A9 antibody. *P < 0.05, **P < 0.01. i, j Representative images and quantification of transwell migration and invasion assay of tumor cells treated with recombinant S100A8/A9 protein dimer. Scale bars, 60 µm. **P < 0.01. k, l Representative images and quantification of colony formation assay of tumor cells treated with recombinant S100A8/A9 protein dimer. **P < 0.01. m Lung-derived metastatic nodules were shown by whole-lung bright, fluorescent and H&E images. Scale bars, 1 mm. n Quantification of lung-derived metastatic nodules. *P < 0.05, **P < 0.01.

Fig 4: S100A8 increases autophagy in BCL cells. (A) Western blotting for LC3-II and P62 in BCL cells was performed following S100A8 interference. (B) Representative images of LC3-positive puncta in BCL cells following S100A8 interference. (C) Transmission electron microscopy of BCL cells following S100A8 interference. Autolysosomes are indicated by arrowheads. Scale bars, 5 µm. (D) Quantification of detectable autolysosomes in BCL cells following S100A8 interference. **P<0.01 and *P<0.05. BCL, B-cell lymphoma; LC3, microtubule-associated protein 1A/1B-light chain 3.

Fig 5: RNA Sequencing analyses revealed differences in innate and adaptive immune responses between sebaceous gland poor (SGP) and sebaceous gland rich (SGR) skin regions (A) Heat map was created by analyzing genes showing significantly different expression (p < 0.05) between SGR (n = 6) and SGP (n = 7) skin. By using StrandNGS software the two sample groups could be distinguished unambiguously based on the level of difference between the gene expression profiles of samples. On the heatmap, this can be seen based on the color scheme of the samples (red: higher expression, blue: lower expression). (B) Principal component analysis (PCA) of RNASeq data (also generated by StrandNGS) showing all samples. On PCA figure each dots represent one skin sample, the color of dots indicate sample type (red: SGP and blue: SGR) and the distance between the dots shows the level of difference between the gene expression profiles of samples. (C) On the basis of Ingenuity Pathway Analysis (IPA), when analyzing genes with significantly different expression between SGP and SGR skin, the first (i.e., the most significant) pathway exclusively related to the skin immune system (SIS) was the IL-17 related one. Genes marked with purple color were present in our gene list subjected to pathway analysis. (D) Regulatory IPA analysis identified three highly overlapping, SIS-related Regulator Effect Networks, in which both upstream regulators and downstream cellular responses were identified in relation to certain gene panels. These networks were linked to immune signaling processes and pathways, which also contained IL-17 related molecules, such as CCL2, S100A8, and S100A9.