Journal: Theranostics

Article Title: Proteomes from AMPK-inhibited peripheral blood mononuclear cells suppress the progression of breast cancer and bone metastasis.

doi: 10.7150/thno.80294

Figure Lengend Snippet: Figure 1. Suppression of tumorigenic behaviors of breast cancer cells by CMLym-Dor. CN = control, CW = CW008 (PKA activator), Dor = Dorsomorphin (AMPK inhibitor), AICAR (AMPK activator), PBMC = peripheral blood mononuclear cells, and CM = conditioned medium. The double asterisks indicate p < 0.01. (A) Suppression of MTT-based viability of MDA-MB-231, MDA-MB-436, and MCF7 breast cancer cells by CMLym-CW and CMLym-Dor, respectively. (B&C) Suppression of scratch-based migration, and transwell invasion of MDA-MB-231 cells by CMLym-Dor. (D) Reduction in p-Src, Snail, and PDL1, together with the elevation in c-Cas3 (cleaved caspase 3) in MDA-MB-231 cells by CMLym-Dor. (E&F) Suppression of MTT-based viability, and transwell invasion of MDA-MB-231 cells by CMPBMC-Dor, which was derived from peripheral blood mononuclear cells with Dorsomorphin.

Article Snippet: Recombinant proteins for ENO1 (700 ng/mL, MBS2009113, MyBioSource, San Diego, CA, USA) and MSN (700 ng/mL, MBS2031729, MyBioSource) were given to MDA-MB-231 cells, and cells were incubated for 24 h. An AMPK inhibitor (Dorsomorphin, 50 μM, #3093, Tocris, Minneapolis, MN, USA) [38], an AMPK activator (AICAR, 50 μM, #2840, Tocris) [39], an activator of PKA signaling (CW008, 20 μM, #5495, Tocris) [40], and an activator of PI3K/Akt (YS49, 10 μM, #HY-15477, MedChem Express, NJ, USA) were applied to the cells for 24 h. Taxol (0.5 nM ~ 5 μM, #1097, Tocris) [41] was applied to breast cancer cells for 48 h. Preparation of CM.

Techniques: Control, Migration, Derivative Assay

Journal: Molecular Metabolism

Article Title: The regulator of G-protein signaling RGS16 promotes insulin secretion and β-cell proliferation in rodent and human islets

doi: 10.1016/j.molmet.2016.08.010

Figure Lengend Snippet: RGS16 knockdown inhibits insulin secretion and is sensitive to PTX and SSTR antagonism . (A–D) Isolated mouse islets were infected with Adv–shRGS16 or Adv-shCTL (scrambled) as a control. Insulin secretion was assessed in 1-h static incubations in response to glucose without or with GLP-1 (100 nM) or carbachol (Carb; 0.5 mM) in control islets ( n = 6–12) (A), after 16 h pretreatment with pertussis toxin (PTX; 100 ng/ml; n = 6) (B), or in the presence of cyclosomatostatin (cSST) at 1 μM ( n = 3) (C) or increasing cSST concentrations ( n = 4) (D). (E) MIN6 cells were infected with Adv–RGS16 or Adv-GFP as a control and insulin secretion was assessed in 1-h static incubations in response to glucose with or without 100 nM SST-14 ( n = 4). Insulin levels are expressed as % of total islet insulin content and are mean ± SEM. * p < 0.05, *** p < 0.001, as compared to the corresponding control.

Article Snippet: Cyclosomatostatin and 8-Bromo-cAMP were from Tocris (Minneapolis, MN, USA).

Techniques: Knockdown, Isolation, Infection, Control

Journal: Molecular metabolism

Article Title: Inhibition of somatostatin enhances the long-term metabolic outcomes of sleeve gastrectomy in mice.

doi: 10.1016/j.molmet.2024.101979

Figure Lengend Snippet: Figure 4: Long-term response of obese mice to SG followed by inhibition of somatostatin signaling. A. Change in weight of SG-cSst mice (purple triangles) and SG-saline mice (black triangles), sham-cSst mice (green circles) and sham-saline mice (black squares). p < 0.01 by treatment and by treatment surgery interaction by 3-way repeated measurement ANOVA. Error bars denote SEM. n ¼ 7,7,9,9. B. Glucose levels following an oral mixed meal tolerance test. Colors as in A. p < 0.01 by treatment 3-way repeated measurement ANOVA. Error bars denote SEM. n ¼ 7,7,9,9. C. Area under the curve (AUC) for the mixed meals tolerance test. D. Fasting plasma insulin levels at the end of the experiment. E,F. Fasting (E) and post-prandial (F) plasma GLP-1 levels at the end of the experiment. G. Post-prandial total cholesterol, HDL-cholesterol, and LDL-cholesterol in the plasma at the end of the experiment. H. Quantification of hepatic steatosis by percent of the area covered by lipid droplets in a hematoxylin and eosin staining. *,**p < 0.05, p < 0.01 by Tukey post-hoc test (A,F,G,H). ##p < 0.01 by surgery in 2-way ANOVA.

Article Snippet: Cyclosomatostatin treatment Cyclosomatostatin (cSst) was purchased from Tocris (Cat. No. 3493) and dissolved to a final concentration of 0.08% EtOH and saline. cSst was injected subcutaneously at 30 mg/kg body weight every evening from 7 days after the surgery until the end of the experiment.

Techniques: Inhibition, Saline, Clinical Proteomics, Staining

Journal: Cancers

Article Title: Conversion of Osteoclasts into Bone-Protective, Tumor-Suppressing Cells

doi: 10.3390/cancers13225593

Figure Lengend Snippet: Conversion of RAW264.7 pre-osteoclasts into iTS cells by the treatment with BML284. CN = control, CM = conditioned medium, RAW = RAW264.7 osteoclasts, MC3T3 = MC3T3 osteoblasts, MSC = mesenchymal stem cells, A5 = MLO-A5 osteocytes, 231 = MDA-MB-231 breast cancer cells, EO = EO771 mammary tumor cells, and 4T1.2 = 4T1.2 mammary tumor cells. ** p < 0.01 vs. CN, while ## p < 0.01 vs. A5 CM. ( A – D ) MTT-based viability of EO771 mammary tumor cells in response to a chemically treated conditioned medium, derived from MLO-A5 osteocytes, MSCs, MC3T3 osteoblasts, and RAW264.7 osteoclasts, respectively. NS = NSC228155 (EGF activator), RC = RCGD423 (JAK/STAT activator), m3 = m-3M3FBS (phospholipase C activator), CW = CW008 (PKA activator), OA = OAC2 (Oct4 activator), YS = YS49 (PI3K activator), and BM = BML284 (Wnt activator). ( E , F ) Tumor selectivity of the inhibitory action of RAW CM, examined tumor selectivity of the inhibitory action using 3 tumor cell lines (MDA-MB-231 breast cancer cell line using 3 tumor cell lines (MDA-MB-231 breast cancer cell line, EO771 mammary tumor cell line, and 4T1.2 mammary tumor cell line), and KTB6 human breast epithelial cells. ( G ) Reduction in PTHrP in BM CM.

Article Snippet: RAW264.7 pre-osteoclast cells, MC3T3 osteoblasts, MSCs, and MLO-A5 osteocyte-like cells were treated with 0.5 μM of NSC228155 (Cayman, Ann Arbor, MI, USA), 20 μM of RCGD423 (Tocris, Minneapolis, MN, USA), 20 μM of m-3M3FBS (Tocris), 20 μM of CW008 (Tocris), 10 μM of OAC2 (MCE, Monmouth Junction, NJ, USA), 50μM of YS49 (MCE), and 0.2 μM of BML284 (Santa Cruz Biotechnology, Dallas, TX, USA) for 1 day.

Techniques: Control, Derivative Assay