Comparable results were seen in analysis of cervical carcinoma lesions (unpublished data). S, stroma. (3.1 MB TIF) pmed.0050019.sg002.tif (3.0M) GUID:?5D128178-A860-4754-942E-F474873E4347 Physique S3: The Abundance of MMP-9CExpressing Cells, or Other Constituent Cell Types of the Neoplastic Cervix, Is Not Altered by Imatinib Therapy (A) Immunostaining of cells expressing MMP-9 (green) in the cervical transformation zone of HPV/E2 mice. Magnification is usually 200; cell nuclei/DAPI, blue; dotted line marks epithelial-stromal boundary. E, epithelium; S, stroma.(B) Immunostaining of the neoplastic cervix using cell-typeCspecific markers (green) revealed no differences in abundance following treatment with imatinib. The cell-type markers were F4/80, macrophages; CD45, leukocytes; mast cell tryptase, mast cells; CD69, NK cells; and CD11c, dendritic cells) Magnification is usually 400; cell nuclei/DAPI, blue. Quantifications were performed using five mice per treatment group. (14 MB TIF) pmed.0050019.sg003.tif (14M) GUID:?36D8AA1C-42C1-4C04-98CD-C206C49BAE5F Physique S4: FGF-2 Is Produced by CAFs Expressing PDGF Receptor- and Vimentin Immunostaining of the stromal compartment of the neoplastic cervix for FGF-2 (red) and for cell type specific markers (green). The markers used to identify the particular cell types are PDGF receptor- for fibroblasts, vimentin for CAFs; CD31 for endothelial cells; F4/80 for macrophages; CD45 for leukocytes; mast cell tryptase for mast cells; CD69 for NK cells; and CD11c for dendritic cells). Magnification is usually 400; cell nuclei/DAPI, blue.(21 MB TIF) pmed.0050019.sg004.tif (21M) GUID:?1B81C449-3799-44CE-B387-082AD3AF3680 Figure S5: Induction of FGF-2 by PDGF in Cultured Fibroblasts FGF-2 transcription was assessed following stimulation of NIH-3T3 mouse fibroblasts with PDGF-AA (100 ng/ml for 6 h in 37 C) T16Ainh-A01 or PDGF-BB (100 ng/ml for 6 h in 37 C). The analysis revealed that fibroblasts up-regulate expression of FGF-2 in response to PDGF. Expression of the housekeeping gene GAPDH was used as a control.(418 KB TIF) pmed.0050019.sg005.tif (418K) GUID:?8AA0F9F5-120E-4C3F-A2DD-6149326A2423 Figure S6: Stromal Cell Density Is Not Altered T16Ainh-A01 in the Neoplastic Cervix following Treatment with Imatinib Quantification of stromal cell density in the stroma of the transformation zone of the cervixes from groups of Rabbit polyclonal to INPP5A five mice treated, or not, with imatinib for 2 wk.(405 KB TIF) pmed.0050019.sg006.tif (406K) GUID:?54B3FDCA-0A8A-4DB2-8B9A-46E451F04F02 Physique S7: PDGF Receptors and FGF-2 Are Expressed in the Stromal Cell Compartment of an Extended Set of Cervical Cancers Representative images of immunohistochemical stainings obtained from the Human Protein Atlas project (http://www.proteinatlas.org/) demonstrate stromal expression of FGF-2 and PDGF receptors.(12 MB TIF) pmed.0050019.sg007.tif (12M) GUID:?F02DB01C-C457-45B8-8DB4-94CA5375D0DB Physique S8: Lack of PDGF Receptor Expression or Functionality on Human Cervical Cell Lines (A) PCR analysis of expression of PDGFR- and PDGFR- by human cervical cancer cell lines (SiHa, C33-A, HeLa) demonstrated that this PDGF-receptor genes were not transcribed. Normal human fibroblasts (NF) were used as a positive control.(B) Western blot analysis of immunoprecipitated PDGFR-  using cell lysates from human cervical cancer cell lines stimulated or not with PDGF-CC demonstrated that PDGFR- was not expressed. Normal human fibroblasts (NF) were used as a control. (C) In vitro analysis of the growth rate of the cervical cancer cell line HeLa produced in the presence or absence of the PDGF-receptor inhibitor imatinib showed that the growth rate of HeLa cells is not altered by the presence of 4.4 M imatinib, corresponding to the peak plasma concentration of imatinib delivered to patients at the standard dose of 400 mg/day. Porcine aortic endothelial (PAE) cells transfected with the PDGFR- were used as a positive control for T16Ainh-A01 the action of imatinib. (3.0 MB TIF) pmed.0050019.sg008.tif (2.9M) GUID:?BCB19D2C-E555-4B2C-9A72-7C8EFFCAC483 Table S1: Expression Level of Growth Factors in the Normal and Neoplastic Cervix Total RNA was extracted from the cervixes of 5-mo-old FVB/n mice treated with estrogen (N/E2), from 3-mo-old HPV/E2 mice (CIN3), or from 5-mo-old HPV/E2 mice (SCC). A pool consisting of five mice from each group was assessed for gene expression using quantitative RT-PCR. The data shown represent the mean from two individual experiments, and are depicted as percent expression of the reference gene L19.(29 KB DOC) pmed.0050019.st001.doc (30K) GUID:?B89DB1D4-C02B-434F-B56C-87C02EEE4169 Table S2: Expression of FGF-2 and PDGF-Receptors in Cervical Cancer Data obtained from meta-analysis of immunohistochemical stainings T16Ainh-A01 performed by the Human Protein Atlas project (http://www.proteinatlas.org). Images were scored for expression in the tumor stroma or neoplastic compartment following careful examination of each specimen displayed on the Web site.(30 KB DOC) pmed.0050019.st002.doc (31K) GUID:?23375297-06F0-465B-BD8B-CF62FBBF6E29 Abstract Background Important support functions, including promotion of tumor growth, angiogenesis, and invasion, have been attributed to the different cell types populating the tumor stroma, i.e., endothelial cells, cancer-associated fibroblasts, pericytes,.
Comparable results were seen in analysis of cervical carcinoma lesions (unpublished data)
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