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. 2018 Feb 26;8(2):24. doi: 10.1038/s41408-018-0063-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Immune histo-chemical staining (brown) of PN-1 in spleen and lymph nodes from normals or infiltrated with DLBCL cells. Blue stain represents haemotoxylin nuclear staining. b Quantitative real-time PCR for the expression of MMP9 and PN1 in lymph nodes hyperplasia (LN) and DLBCL. N = 6, ***P < 0.001. Student's t test was performed for statistical significance. c DAB staining of PN-1 (brown, panel i) and relative expression score (ii) in controls with a normal bone marrow (N = 9), persons with lymphoma with an involved (N = 7) or uninvolved bone marrow (N = 6). One-way ANOVA performed for statistical significance. d Proliferation of Raji cells (2 × 10⁶) was analyzed by using CCK-8 assays at 24, 48, 72, and 96 h after transfection with 2 μg of pcDNA3-PN1 or control plasmid (i) or followed by treatment of 40 pmol siRNA PN1 (siPN1) or control scrambled siRNA (siNEG) (N = 3; t test *P < 0.05; **P < 0.01; ***P < 0.001); e Apoptosis analysis was determined by Annexin-PI assay (i) and immune-blotting of cleavage of caspase-3 (15-kDa active band) (ii) in Raji cells transfected with 2 μg of pcDNA3-PN1 or control plasmid (N = 4; t test **P < 0.01); f Raji cells transfected with siRNA PN1 (siPN1) or control siRNA (siNEG) were assayed for apoptosis (N = 3; t test *P < 0.05); g Subcutaneous tumor volumes from Balb/c mice implanted with A20 cells (1 × 10⁶) mixed with Matrigel with or without PN-1 recombinant protein (10 μM). Subcutaneous tumor growth curve (i) (N = 5; t test, **P < 0.01; ***P < 0.001) and tumor volumes (ii); immune-blotting detecting caspase-3 cleavage (iii); h A20 xenografts with or without pre-treatment with PN-1 (10 μM) recombinant protein, DAB-stained (brown) for caspase-3 and Ki67. Blue stain represents hematoxylin nuclear staining. (i) in situ apoptotic assay using TUNEL. Positive cells (red stain) were counted from three microscopic fields and plotted (N = 4, t test, **P < 0.01). Nuclei are blue stained