Journal: Nature medicine

Article Title: The Cyclophilin A-CD147 complex promotes bone marrow colonization of B-cell malignancies: implications for therapy

doi: 10.1038/nm.3867

Figure Lengend Snippet: Transwell migration assays of MM1S-luc cells incubated under different conditions: ( a ) increased concentrations of recombinant eCyPA.; ( b ) medium alone or BMEC-60 cells lentivirally transduced with Control-shRNA or shRNAs against CyPA (CyPA-shRNA). Migration data was normalized based on data of medium alone. Results are means ± SD for assays performed in triplicate. Statistical significance of differences between groups was determined by unpaired Student's t-test. ( c ) Immunoblot of total protein extracts from H929 and MM1S cells incubated in the absence (-) or presence (+) of eCyPA at 50ng per ml. Xenogen data ( d ), time course ( e ), and histologic analysis ( f ) of MM1S-luc cell growth within scaffolds coated with BMEC-60 lentivirally transduced with ControlshRNAs or CyPA-shRNA. Bars: Top and midle 20μm, Bottom 100μm. The results of one representative of three independent experiment is shown Xenogen data ( g ), time course ( h ), and histologic analysis ( i ) of cell growth of MM1S-luc transduced with Control-shRNA or CD147-shRNA within empty scaffolds or scaffolds coated with BMEC-60 cells. Bar: 100μm. Statistical analyses of tumor burden were done using factorial analysis in SPSS 13.0. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).

Article Snippet: Anti-human primary antibodies included: BCL9 (2D4, Abnova, 1:1000), CyPA (ERPR7511, Abcam, 1:1000), CD147 (13287, Cell Signaling, 1:1000), pSTAT3 (9131, Cell Signaling, 1:1000), pERK (4370, Cell Signaling, 1:1000), PARP (9542, Cell Signaling, 1:1000), pAKT (9271, Cell Signaling, 1:1000), Active-β-catenin (05-665, Millipore, 1:1000), Factor VIII (A0082, DAKO, 1:1000), MMP-9 (MAB911, R&D, 1:1000), CyPB (MAB5410, R&D, 1:500), and horseradish peroxidase(HRP)-conjugated actin (C-11, Santa Cruz, 1:200).

Techniques: Migration, Incubation, Recombinant, Transduction, shRNA, Western Blot

Journal: Nature medicine

Article Title: The Cyclophilin A-CD147 complex promotes bone marrow colonization of B-cell malignancies: implications for therapy

doi: 10.1038/nm.3867

Figure Lengend Snippet: ( a ) Time course of Xenogen imaging of MM1S-luc cell growth in scaffolds implanted inCB17.Cg-PrkdcscidLystbg-J-Crl mice and treated with local injections of isotype control or anti-CD147 Abs. Xenogen data ( b ) of MM1S-luc cell growth within scaffolds coated with BMEC-60 cells and implanted subcutaneously in CB17.Cg- Prkdc scid Lyst bg-J -Crl mice. Groups of 4 mice were subsequently treated with either isotype Ab or anti-CD147 Ab, and tumor growth within the scaffolds was evaluated by Xenogen imaging every five days. ( c ) Left panel, Immunofluorescence analysis of CD147 expression in MM plasma cells from BM (top) and PB (bottom) from one person with MM (Case 1). Right panel, Immunofluorescence analysis of CD147 expression in normal plasma cells from BM (top) and lymph node (LN) (bottom) in two different normal donors (Case 3 and 4). Bars: 5μm. ( d ) Proposed model of BM homing of MM cells based on eCyPA secreted by BMECs and on CD147 expression by MM cells. Statistical analysis of tumor burden were done using factorial analysis in SPSS 13.0. (*P<0.05, **P<0.01).

Article Snippet: Anti-human primary antibodies included: BCL9 (2D4, Abnova, 1:1000), CyPA (ERPR7511, Abcam, 1:1000), CD147 (13287, Cell Signaling, 1:1000), pSTAT3 (9131, Cell Signaling, 1:1000), pERK (4370, Cell Signaling, 1:1000), PARP (9542, Cell Signaling, 1:1000), pAKT (9271, Cell Signaling, 1:1000), Active-β-catenin (05-665, Millipore, 1:1000), Factor VIII (A0082, DAKO, 1:1000), MMP-9 (MAB911, R&D, 1:1000), CyPB (MAB5410, R&D, 1:500), and horseradish peroxidase(HRP)-conjugated actin (C-11, Santa Cruz, 1:200).

Techniques: Imaging, Immunofluorescence, Expressing