Journal: Journal for Immunotherapy of Cancer

Article Title: Synergizing sunitinib and radiofrequency ablation to treat hepatocellular cancer by triggering the antitumor immune response

doi: 10.1136/jitc-2020-001038

Figure Lengend Snippet: Sunitinib (SU) treatment repressed RFA–induced PD-1 upregulation via HGF signaling. Mice with size-matched tumors were randomly distributed into four groups and received the indicated treatments as described in . Two weeks post-RFA, tumors were harvested from each mouse and used for the following studies. (A) The gene expression of HGF, c-Met, HIF-1, and IL-6 in the tumors. Total RNAs were extracted from part of tumors in the different groups of mice. qPCR was used to detect the mRNA expression of HGF, c-Met, HIF-1, and IL-6. n=4, *p<0.05, **p<0.01, ***p<0.001, error bars represent means±SD. (B) HGF protein expression in the tumors. Tumor lysate was prepared and used to detect HGF protein expression by western blotting. The protein quantification assay was conducted with ImageJ. n=4, ***p<0.001, error bars represent means±SD. (C) Expression of PD-1 in CD4 + and CD8 + T cells in response to the different stimulation. The splenocytes from wild-type mice were stimulated with the basal level anti-CD3 Ab and the tumor lysate from the mice with the different treatments. Flow cytometry was used to detect PD-1 expression in CD4 + and CD8 + T cells. iHGF, HGF inhibitor; rHGF, recombinant HGF; TL-CTR, tumor lysate from the control mice without treatment; TL-RFA, tumor lysate from the mice with RFA monotherapy; TL-RFA+SU, tumor lysate from the mice with combined treatment with SU and RFA. (D) Average frequencies of PD-1 + CD4 + T cells and PD-1 + CD8 + T cells. n=3, **p<0.01, ***p<0.001, error bars represent means±SD. Statistical analysis was performed by Student t-test.

Article Snippet: These cells were stimulated with tumor lysates prepared from tumor-bearing mice with different treatments in the presence or absence of recombinant HGF protein (10 µg/mL) (Bio Legend, San Diego, CA), or neutralized anti-HGF antibody (10 µg/mL) (Sino Biological, Beijing, China) for 72 hours, then harvested for the following studies.

Techniques: Expressing, Western Blot, Flow Cytometry, Recombinant

Journal: Arthritis and rheumatism

Article Title: Impairment of the antifibrotic effect of hepatocyte growth factor in lung fibroblasts from African Americans: possible role in systemic sclerosis.

doi: 10.1002/art.22713

Figure Lengend Snippet: Figure 2. Levels of cytokines in bronchoalveolar lavage (BAL) fluid from Caucasian and African American patients with systemic sclerosis (SSc) and normal controls. Cytokine levels were significantly elevated in BAL fluid from African American control subjects as compared with Caucasian control subjects. A, Levels of hepatocyte growth factor (HGF), insulin-like growth factor binding protein 3 (IGFBP-3), osteoprotegerin (OPG), stem cell factor (SCF), thrombopoietin (TPO), and vascular endothelial growth factor (VEGF) were measured by cytokine array analysis. Array images were scanned and analyzed with National Institutes of Health Image software. Positive controls (6 per membrane) were used to normalize the results from the different membranes being compared. Values are the mean and SEM of 24 separate arrays of BAL fluid samples from controls and 40 separate arrays from SSc patients. Variation between 2 identical cytokine spots ranged from 0% to 10% in duplicate experiments. B, Effect of selected cytokines on type I collagen expression in lung fibroblasts from Caucasian and African American normal subjects and SSc patients. Confluent cultures of lung fibroblasts were serum-starved for 24 hours and then incubated for 72 hours with recombinant transforming growth factor (TGF; 5 ng/ml), which was used as a positive control in studies of normal lung fibroblasts, or with HGF, IGFBP-3, OPG, SCF, TPO, or VEGF (50 ng/ml each). Serum-free medium (SFM) was used as a basal level for type I collagen. Cells were collected with lysis buffer and analyzed by Western blotting using anti–type I collagen antibody. Anti–-actin antibody was used as the sample loading control. Shown are representative immunoblots from 3 independent experiments.

Article Snippet: Confluent cultures of lung fibroblasts were serum-starved for 24 hours, followed by incubation for 72 hours with recombinant transforming growth factor (TGF ; 5 ng/ml) and/or HGF (50 ng/ml or concentrations indicated below), insulin-like growth factor binding protein 3 (IGFBP-3), osteoprotegerin (OPG), stem cell factor (SCF), thrombopoietin (TPO), and vascular endothelial growth factor (VEGF), with or without neutralizing anti-human HGF receptor (c-Met) antibody (all purchased from R&D Systems).

Techniques: Control, Binding Assay, Software, Membrane, Expressing, Incubation, Recombinant, Positive Control, Lysis, Western Blot

Journal: Arthritis and rheumatism

Article Title: Impairment of the antifibrotic effect of hepatocyte growth factor in lung fibroblasts from African Americans: possible role in systemic sclerosis.

doi: 10.1002/art.22713

Figure Lengend Snippet: Figure 5. Modulation of the effects of hepatocyte growth factor (HGF) on connective tissue growth factor (CTGF) expression and type I collagen accumulation by anti–c-Met antibody treatment of lung fibroblasts from Caucasian patients with systemic sclerosis (SSc) and normal controls. Anti–c-Met antibody (Ab) diminished the effects of HGF on CTGF expression and collagen accumulation. Serum-starved lung fibroblasts were incubated for 72 hours with medium containing anti–c-Met antibody (2 g/ml), with or without HGF (50 ng/ml). Normal lung fibroblasts were left untreated or were additionally treated with transforming growth factor (TGF; 5 ng/ml). Anti–- actin antibody was used as the sample loading control. Shown are representative immunoblots from 3 independent experiments.

Article Snippet: Confluent cultures of lung fibroblasts were serum-starved for 24 hours, followed by incubation for 72 hours with recombinant transforming growth factor (TGF ; 5 ng/ml) and/or HGF (50 ng/ml or concentrations indicated below), insulin-like growth factor binding protein 3 (IGFBP-3), osteoprotegerin (OPG), stem cell factor (SCF), thrombopoietin (TPO), and vascular endothelial growth factor (VEGF), with or without neutralizing anti-human HGF receptor (c-Met) antibody (all purchased from R&D Systems).

Techniques: Expressing, Incubation, Control, Western Blot

Journal: Arthritis and rheumatism

Article Title: Impairment of the antifibrotic effect of hepatocyte growth factor in lung fibroblasts from African Americans: possible role in systemic sclerosis.

doi: 10.1002/art.22713

Figure Lengend Snippet: Figure 6. Hepatocyte growth factor (HGF)–induced c-Met receptor phosphorylation in lung fibroblasts from Caucasian and African American patients with systemic sclerosis (SSc). Lung fibroblasts were cultured to confluence on 6-well plates and collected with lysis buffer. Twenty micrograms of protein per sample was resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (4–20% gradient gels) and analyzed by Western blotting using anti–phospho–c-Met (Ph-C-Met) or anti–c-Met polyclonal antibodies (see Patients and Methods for details). Anti–-actin antibody was used as the sample loading control. Shown are representative immunoblots from 3 independent experiments. A, Levels of phosphor- ylated and total c-Met receptor in lung fibroblasts from Caucasian and African American SSc patients. B, Concentration-dependent phosphorylation of c-Met receptor in lung fibroblasts from Caucasian and African American SSc patients treated with HGF for 5 minutes at the indicated concentrations. C, Time course of c-Met receptor phosphorylation in lung fibroblasts from Caucasian and African American SSc patients after treatment with HGF (50 ng/ml) for various periods (left). Western blots from the time course experiments were analyzed by scanning densitometry using National Institutes of Health Image software (right). Values are the mean of 4 samples at each time point.

Article Snippet: Confluent cultures of lung fibroblasts were serum-starved for 24 hours, followed by incubation for 72 hours with recombinant transforming growth factor (TGF ; 5 ng/ml) and/or HGF (50 ng/ml or concentrations indicated below), insulin-like growth factor binding protein 3 (IGFBP-3), osteoprotegerin (OPG), stem cell factor (SCF), thrombopoietin (TPO), and vascular endothelial growth factor (VEGF), with or without neutralizing anti-human HGF receptor (c-Met) antibody (all purchased from R&D Systems).

Techniques: Phospho-proteomics, Cell Culture, Lysis, Polyacrylamide Gel Electrophoresis, Western Blot, Control, Concentration Assay, Software

Journal: American Journal of Translational Research

Article Title: Combination of cetuximab with met inhibitor in control of cetuximab-resistant oral squamous cell carcinoma

doi:

Figure Lengend Snippet: HGF induction of OSCC resistance to cetuximab. A. Western blot. HSC-2 and HSC-3 cells were subjected to protein extraction and showed high protein expression levels of EGFR and MET by western blot. B. The MTT assay. HSC-2 and HSC-3 cells were treated with different doses of cetuximab (0-5 µg/mL) and HGF (20 ng/mL) for 72 h, and the cell proliferation levels were shown by the MTT assay. C. Western blot. HSC-2 and HSC-3 cells were pretreated with control, HGF (20 ng/mL), cetuximab (1 µg/mL), or their combination. D. The MTT assay. HSC-2 cells treated with HGF (20 ng/mL) and the HGF neutralizing antibody or cetuximab (1 µg/mL) for 72 h showed different proliferation abilities by the MTT assay. *P < 0.05.

Article Snippet: In brief, 2 × 10 3 tumor cells/well were seeded into 96-well plates and grown for 24 h and then treated with various concentrations of cetuximab (Merck, Germany), PHA-665752 (Selleck Chemicals, TX, USA), HGF (R&D Systems, Minneapolis, MN, USA), and a goat anti-human HGF neutralizing antibody, normal goat IgG (R&D Systems) for 72 h. At the end of each experiment, 50 μL of MTT solution (2 mg/mL; Sigma Chemicals, St. Louis, MO, USA) was added to each well of the cell culture plate, and the cells were incubated for an additional 2 h. The culture medium was replaced with 100 μL of dimethyl sulfoxide.

Techniques: Western Blot, Protein Extraction, Expressing, MTT Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Hepatocyte Growth Factor: A Microenvironmental Resource for Leukemic Cell Growth

doi: 10.3390/ijms20020292

Figure Lengend Snippet: ( A ) Percentage of estimated CLL cells survival benefit due to conditioned medium or cell-to-cell contact (co-cultures). Depicted bars indicate the percentages of viable CLL cells deprived of relative control percentages in all experimental conditions, using the indicated different cell types (CTR co-cultures: 32.17 ± 31.62; CTR conditioned medium: 63 ± 22). The reported number of cases used ( n ) are color-linked to the relative bars and were used to generate the corresponding SD values. ( B ) Estimated reduction of CLL survival benefit due to addition of c-MET inhibitor or anti-HGF moAb in co-cultures or during exogenous HGF administration. Depicted bars indicate the percentages of viable CLL cells deprived of relative control percentages in all experimental conditions (CTR-HGF:70 ± 9.5: CTR-BMSC:58.8 ± 6.5; CTR-MG63:47.5 ± 22.9) using the indicated different cell types. The reported number of cases used (n) are color-linked to the relative bars and were used to generate the corresponding SD values. SU: SU11274, αHGF:HGF neutralizing moAb L2G7. Statistical significance (* p ≤ 0.05; **: 0.01 ≤ p < 0.001; ***: p ≤ 0.001; Student t -test) is referred to each culture condition in comparison to relative controls (CLL cells in medium only, here not shown as above described) and the present Figure is a different representation with minor modifications of previously published data reported by Giannoni P et al. in .

Article Snippet: We demonstrated that the c-MET tyrosine kinase inhibitor SU11274 or the neutralizing anti-HGF moAb L2G7 (a kind gift from Galaxy Biotech, Sunnyvale, CA 94089, USA) counteracted increased CLL cells viability induced by HGF treatment or after co-culturing leukemic cells with specific stromal cells (BMSC, trabecular bone cells, the osteoblasto-like cell line MG63 [ ] and ).

Techniques: Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Hepatocyte Growth Factor: A Microenvironmental Resource for Leukemic Cell Growth

doi: 10.3390/ijms20020292

Figure Lengend Snippet: Hypothetical model of associated therapies targeting CLL-support from microenvironment and BCR signaling: HGF released in paracrine and autocrine manner within the CLL microenvironment maintains viability of accessory cells (stromal, osteoblasts cancer associated fibroblasts, and nurse-like cells) which in turn support viability of leukemic cells. The usage of anti-HGF moAb (i.e., L2G7, AMG102), should block CLL support by cells of the microenvironment. On the other side inhibitors of the tyrosine kinase c-MET (i.e., SU11274, Crizotinib, Tivantinib), c-MET siRNA transfected exosomes or a c-MET directed aptamer, in association with inhibitors of tyrosine kinases triggered by BCR (PI3Kd, SYK, BTK), may directly interfere with survival and expansion of the leukemic B cell clone promoted by the inner cell signaling pathways.

Article Snippet: We demonstrated that the c-MET tyrosine kinase inhibitor SU11274 or the neutralizing anti-HGF moAb L2G7 (a kind gift from Galaxy Biotech, Sunnyvale, CA 94089, USA) counteracted increased CLL cells viability induced by HGF treatment or after co-culturing leukemic cells with specific stromal cells (BMSC, trabecular bone cells, the osteoblasto-like cell line MG63 [ ] and ).

Techniques: Blocking Assay, Transfection, Protein-Protein interactions

Journal: Frontiers in Oncology

Article Title: Increased RAB31 Expression in Cancer-Associated Fibroblasts Promotes Colon Cancer Progression Through HGF-MET Signaling

doi: 10.3389/fonc.2020.01747

Figure Lengend Snippet: HGF/MET signaling blockade abolishes colon cancer cell migration mediated by RAB31 in CAFs. (A) RKO migration measured by Transwell assay at 50 h. The lower chamber contained primary culture medium supplemented with 6% FBS for control or CAF derived CM. (B) The migration of RKO was significantly enhanced by vector control (EGFP) CAF CM, and further enhanced by RAB31 overexpressing CAF CM measured by Tranwell assay. Knockdown of RAB31 in CAF by lentivirus significantly reduced the migration of RKO. **** P < 0.0001 vs. CAF23 EGFP CM. (C,D) Wound healing assay of RKO cells incubated with CM from vector control CAFs or RAB31 knockdown CAFs. *** P < 0.001 vs. RAB31 KD CM. (E) Migration of RKO cells treated with RAB31 overexpressing CAF CM or EGFP CAF CM (vector control) in the presence of MET inhibitors Crizotinib (100 nM) or Su11274 (5 μM) measured by Transwell assay at 45 h. (F) Blockade of MET signaling significantly inhibited the CAF CM mediated RKO migration and ameliorated the promigratory effect of RAB31 overexpression. **** P < 0.0001 vs. CAF23 RAB31 CM control; ### P < 0.001, # P < 0.05 vs. CAF23 EGFP CM. (G) WB indicated that treatment of RKO with CAF23 EGFP CM increased the phosphorylation of MET, which was blocked by MET inhibitors or by MET knockdown in RKO. (H,I) Neutralizing HGF in CAF25 EGFP CM and CAF25 RAB31 CM inhibited migration of RKO cells by Transwell assay at 45 h in a dose dependant manner and abolished the difference of the two CMs at 5 μg/ml. #### P < 0.0001, # P < 0.05 vs. CAF23 EGFP CM. (J) WB results showed that the increased MET phosphorylation in RKO induced by CM from RAB31 overexpressing CAFs was diminished upon preincubation with anti-HGF antibody at 5 μg/ml. (D,F,G,I) Data shown as mean ± s.d. Statistical significance was determined by Student's t -test.

Article Snippet: For HGF neutralization experiments, the anti-HGF polyclonal antibody (AF-294-NA, R&D Systems, Minneapolis, MNB, USA) was added to the mixed medium and incubated at 37°C for 1 h before adding to the lower chamber.

Techniques: Migration, Transwell Assay, Control, Derivative Assay, Plasmid Preparation, Knockdown, Wound Healing Assay, Incubation, Over Expression, Phospho-proteomics

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 2. Conditioned media from BMPC regulates miR-155 expression and fibrogenic response in mouse cardiac fibroblasts in vitro. (A) conditioned media (CM) from BMPC reduced miR-155 expression (A) and mRNA expression of fibrogenesis markers like collagen type1 alpha 1(col1A1) (B), col3A1 (C) and a-SMA (D) in mouse cardiac fibroblasts induced under diabetic condition [TGF-beta (10 mg/mL+ high glucose (25 mM glucose)]. Neutralizing antibodies against HGF (HGF-Ab) reversed this effect as compared to IgG treated cells. *P,0.01 versus control; #P,0.05 versus without CM; $P,0.05 versus BMPC conditioned media with IgG. E. Protein expression of collagen I and a-SMA in cardiac fibroblasts with miR-155 over-expression was determined by Western blotting. b-actin served as loading controls. C indicates control condition (5.5 mM glucose); BMPC-CM, conditioned media from BMPC; control supernatant, control media; HGF-Ab, antibodies against hepatocyte growth factor; Col1A1, collagen type 1 alpha 1; Col1A1, collagen type 3 alpha 1; a-SMA, alpha smooth muscle actin. doi:10.1371/journal.pone.0060161.g002

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Expressing, In Vitro, Control, Over Expression, Western Blot

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 3. Expression of miR-155 in infarcted hearts (day 3) of db/db mice receiving BMPC intramyocardial. BMPC therapy marginally (although not significantly) decreased MI-induced miR-155 expression (measured by qRT-PCR) in comparison with saline-treated group. Administration of neutralizing antibodies against HGF (HGF-Ab) significantly elevated miR-155 levels as compared to IgG treated group. *P,0.05 versus sham; #P,0.01 versus IgG treated mice. HGF-Ab indicates antibodies against hepatocyte growth factor; Col1A1, collagen type 1 alpha 1; Col1A1, collagen type 3 alpha 1; a-SMA, alpha smooth muscle actin; BMPC, bone marrow-derived progenitor cell. BMPC, bone marrow-derived progenitor cell; MI, myocardial infarction. *P,0.01 vs sham; #P,0.05 vs IgG treated group. doi:10.1371/journal.pone.0060161.g003

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Expressing, Quantitative RT-PCR, Comparison, Saline, Derivative Assay

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 4. Cardiac fibrosis in db/db mice receiving BMPC therapy. Fibrosis area was determined at 28 days after MI by Sirius red staining. Collagen stains deep red and normal tissue stains yellowish. BMPC therapy decreased % fibrosis in comparison with saline-treated group. Administration of neutralizing antibodies against HGF (HGF-Ab) significantly aggravated fibrosis as compared to IgG treated group. Bar graph represents quantification of percent fibrosis area (percentage of LV area). HGF-Ab indicates antibodies against hepatocyte growth factor; BMPC, bone marrow-derived progenitor cell. BMPC, bone marrow-derived progenitor cell. *P,0.05 vs saline group; #P,0.05 versus IgG treated group. doi:10.1371/journal.pone.0060161.g004

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Staining, Comparison, Saline, Derivative Assay

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 5. Echocardiographic analysis of LV function in db/db mice receiving BMPC therapy after 28 days post-MI. Ejection fraction (EF) increased following BMPC therapy (improvement in cardiac function) in comparison with saline-treated group. Administration of neutralizing antibodies against HGF (HGF-Ab) worsened LV function as compared to IgG treated group. *P,0.05 vs saline group; #P,0.05 versus IgG treated group. HGF-Ab indicates antibodies against hepatocyte growth factor; BMPC, bone marrow-derived progenitor cell. BMPC, bone marrow-derived progenitor cell. doi:10.1371/journal.pone.0060161.g005

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Comparison, Saline, Derivative Assay

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 6. Expression of fibrotic markers (at 28 day post-MI) in infarcted hearts of db/db mice receiving BMPC intramyocardial. BMPC therapy reduced MI-induced mRNA expression (measured by qRT-PCR) of Col1A1, Col3A1 and a-SMA in comparison with saline-treated group. Administration of neutralizing antibodies against HGF (HGF-Ab) significantly aggravated fibrotic response as compared to IgG treated group. #P,0.05 versus IgG treated group. HGF-Ab indicates antibodies against hepatocyte growth factor; Col1A1, collagen type 1 alpha 1; Col1A1, collagen type 3 alpha 1; a-SMA, alpha smooth muscle actin; BMPC, bone marrow-derived progenitor cell. BMPC, bone marrow-derived progenitor cell. doi:10.1371/journal.pone.0060161.g006

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Expressing, Quantitative RT-PCR, Comparison, Saline, Derivative Assay

Journal: PloS one

Article Title: Bone marrow progenitor cell therapy-mediated paracrine regulation of cardiac miRNA-155 modulates fibrotic response in diabetic hearts.

doi: 10.1371/journal.pone.0060161

Figure Lengend Snippet: Figure 9. Proposed mechanisms of the paracrine regulation of cardiac fibrosis by transplanted BMPCs. BMPCs release HGF, which inhibits miR-155 leading to de-repression of inhibitors of TGF- b signaling, Ski/SnoN (possible targets of miR-155), thereby inhibiting cardiac fibrosis. HGF indicates hepatocyte growth factor; BMPC, bone marrow-derived progenitor cell. MI indicates myocardial infarction; BMPC, bone marrow-derived progenitor cell; HGF, hepatocyte growth factor; TGF-b, TGF-beta; Ski, Sloan-Kettering Institute proto-oncogene; SnoN, Ski-related novel gene, non-Alu-containing. doi:10.1371/journal.pone.0060161.g009

Article Snippet: In vivo Neutralization of HGF by an Anti-HGF Antibody or Administration of Recombinant Mouse HGF Protein To determine if the antifibrosis effect of BMPC treated mice is inhibited by blocking HGF signaling, a subset of mice that underwent MI surgery and BMPC transplantation were injected intraperitoneally with neutralizing goat anti-HGF antibodies (25 mg/kg b.w. per day for seven days; R&D Systems, Minneapolis, MN) or normal goat IgG for 7 days, post-MI.

Techniques: Derivative Assay