Fig. 1: Breast and prostate cancer cells repress Tsp-1 in a paracrine manner via PRSS2. | Nature Communications

Fig. 1: Breast and prostate cancer cells repress Tsp-1 in a paracrine manner via PRSS2.

From: PRSS2 remodels the tumor microenvironment via repression of Tsp1 to stimulate tumor growth and progression

Fig. 1

A Immunohistochemistry (IHC) of thrombospondin-1 (Tsp-1) expression in tumors formed in the prostate gland of SCID mice by PC3 and PC3M-LN4 prostate cancer cells (scale bar = 100 μm) (n = 8 per group). B Schematic diagram of the proteomic screening and identification of Tsp-1 repressing protein in PC3M-LN4 conditioned media. C ELISA of Tsp-1 expression (normalized to total protein levels) in PC3M-LN4 fractions eluted from a Heparin-sepharose-Cu2+ column (n = 3). D Western blot of PRSS2 and actin protein levels in PC3 and PC3M-LN4 prostate cancer cells (n = 3). E Western blot of PRSS2 and actin protein levels in MDA-MB-231 (231), MDA-MB-231-LM2 (LM2), MCF7 and SUM159 breast cancer cells (n = 3). F Tsp-1 and actin levels in MRC5 fibroblasts and primary human peripheral blood mononuclear cells (PBMCs) that were untreated (–) or treated with recombinant human PRSS2 ( + ) (n = 3). G Plot of Tsp-1 mRNA fold change, measured by real-time quantitative RT-PCR in WI38 cells treated with conditioned media of 293T cells engineered to overexpress PRSS2 (n = 3; P value calculated via two-sided Student’s t test). H Upper panel: Western blot of PRSS2 in SUM159 cells that were untransfected (C), transfected with empty pLKO.1 vector (V), or pLKO.1 vector expressing three independent shRNA sequences directed against PRSS2 (sh1, sh2, sh3). Lower panel: Western blot of Tsp-1 and actin in WI38 cells that were untreated (–) or treated with conditioned media from SUM159 cells with empty vector (V) or PRSS2 shRNA (n = 3). I Western blot of Myc, PRSS2 and actin in SUM159 cells transfected with vector control (V) or three shRNA sequences directed against c-Myc (sh1, sh2 and sh3) (n = 3). J Dot plot of fold change of Myc and PRSS2 in SUM159 cells transfected with vector control (V) or three shRNA sequences directed against c-Myc (sh1, sh2, and sh3 (n = 3). K Dot plot of fold change of Myc and PRSS2 in PC3M-LN4 cells transfected with vector control (V) or three shRNA sequences directed against c-Myc (sh1, sh2, and sh3) (n = 3).

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