Journal: Nutrients

Article Title: Manuka Honey Inhibits Human Breast Cancer Progression in Preclinical Models

doi: 10.3390/nu16142369

Figure Lengend Snippet: Manuka honey activates AMPK signaling and inhibits mTOR and STAT3 downstream signaling. ( A ) MCF7 cells were treated with increasing concentrations of Manuka honey (0–5%) and 5% Mesquite honey as control. After 24 h, cells were lysed and immunoblotted with specific antibodies. ( B ) MCF7 cells were treated with increasing concentrations of Manuka honey 0.3–5% ( w / v ). After 24 h, cells were lysed, and whole cell extracts were resolved by PAGE and immunoblotted with specific antibodies.

Article Snippet: MCF7 and MDA-MB-231 BC cells were cultured in 6-well plates at a concentration of 250,000 cells/well for 24 h. The next day, cells were treated with either 2.5% MH, 5.0% MH, 5.0% dextrose, 5.0% Mesquite honey, or 10 μM Camptothecin for 24 and 48 h. Cells were collected with AccutaseTM cell dissociation reagent, washed and stained using an Annexin V-FITC and 7-AAD Dead Cell Apoptosis Detection Kit (R&D Systems), and analyzed using flow cytometry.

Techniques: Control

Journal: Cancer Cell

Article Title: Genomic and Transcriptomic Determinants of Therapy Resistance and Immune Landscape Evolution during Anti-EGFR Treatment in Colorectal Cancer

doi: 10.1016/j.ccell.2019.05.013

Figure Lengend Snippet: Transcriptomic CRC Subtypes and CAFs as Drivers of AR to Cetuximab (A) Transcriptomic subtypes in 13 BL and PD biopsy pairs. TA, transit amplifying; SL, stem-like. (B) Volcano plot showing differential expression of growth factors in 5 cases from (A) undergoing CMS2>4 switches. Significance was assessed by paired t test. (C) Changes in TGF-β and EMT transcriptomic signatures through CMS2>4 switches. (D) Changes in fibroblast abundance through CMS2>4 switches based on MCP-counter analysis. (E) Impact of CAF conditioned medium (CM) on the growth of DiFi (left panel) and LIM1215 (right panel) treated with 50 μg/mL CET for 5 days. (F) Western blot analysis showing CAF CM rescue of pERK in DiFi (left panel) and LIM1215 (right panel) treated with 200 μg/mL CET for 2 h. (G) mRNA expression (normalized counts) of growth factors (GFs) (left panel) and their receptors (right panel) in CAF, DiFi, and LIM1215 cells. (H) Growth assay with 200 μg/mL CET and recombinant GF at a concentration of 20 ng/mL (FGF1/2), 10 ng/mL (TGF-β) and 50 ng/mL (HGF) for 5 days in DiFi (top panel) and LIM1215 (bottom panel). (I) Western blot analysis of pERK with and without recombinant GF treatment in the presence or absence of 200 μg/mL CET in DiFi (top panel) and LIM1215 (bottom panel). (J) Growth assay with CAF CM and combinations of CET, pan-FGFR inhibitor (FGFRi), and MET inhibitor (METi) for 5 days in DiFi (top panel) and LIM1215 (bottom panel). (K) Western blot analysis of pERK after 2 h treatment with CAF CM and combinations of CET, FGFRi, and METi in DiFi (top panel) and LIM1215 (bottom panel). (E, H, and J) All error bars ± SD of six replicates. See also Figure S4 and .

Article Snippet: Recombinant human FGF1 acidic , RnD Systems , Cat# 232-FA-025.

Techniques: Quantitative Proteomics, Western Blot, Expressing, Growth Assay, Recombinant, Concentration Assay

Journal: Cancer Cell

Article Title: Genomic and Transcriptomic Determinants of Therapy Resistance and Immune Landscape Evolution during Anti-EGFR Treatment in Colorectal Cancer

doi: 10.1016/j.ccell.2019.05.013

Figure Lengend Snippet:

Article Snippet: Recombinant human FGF1 acidic , RnD Systems , Cat# 232-FA-025.

Techniques: Control, Recombinant, Sequencing, Software

Journal: JCI insight

Article Title: Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1.

doi: 10.1172/jci.insight.132796

Figure Lengend Snippet: Figure 1. CHIR99021 and/or FGF1 NP treatment: assessment of cardiac function and infarct size in a mouse model of MI. Mice that were treated with intramyocardial injections of different NPs, including CHIR + FGF1-NPs, CHIR- NPs, FGF1-NPs, and empty NPs (nonloaded); MI-only control mice; and sham-operated control mice were subjected to echocardiographic assessments of left ventricular (LV) function (A). Ejection fraction (EF) (B), fractional shorten- ing (FS) (C), end-systolic diameter of the left ventricle (D), and end-diastolic diameter of the left ventricle (E) were assessed before MI induction (pre-S) and on post-MI day 28 (post-S). On day 28 after MI, CHIR + FGF1-NP treatment groups presented significantly greater EF and FS compared with other treatment as well as control groups (B and C) and significantly lower values of systolic/diastolic diameters of the left ventricle. Data are given as means ± SEM. There were 10–12 animals per group. Statistical analysis: 2-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. sham; †P < 0.01 vs. MI; ‡P < 0.05 vs. empty NPs; §P < 0.05 vs. CHIR-NPs, ||P < 0.01 vs. FGF1-NPs. Sirius Red/Fast Green histochemical staining revealing areas of infarcted (red, nonviable) and noninfarcted (green, viable) zones in post-MI day 28 ventricular tissue sections (F). The infarct size was quantified as the ratio of the scar area to the total surface area of the left ventricle and expressed as a percentage, for day 28 samples (G). At day 28, the CHIR + FGF1-NP treatment group showed significant reduction in infarct size compared with other NP treatment groups or the untreated control MI animals. Scale bar: 1 mm (panels in F). Data are given as means ± SEM. There were 10–12 animals per group. Statistical analysis: 1-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. MI; †P < 0.01 vs. empty NP; ‡P < 0.01 vs. CHIR-NP; §P < 0.01 vs. FGF1-NP.

Article Snippet: A double-emulsion (water/oil/water phase) technique for recombinant human FGF1 acidic protein (rhFGF1, aa 16-155 (Novus Biologicals, Inc.) and a single-emulsion (oil/water phase) technique for CHIR99021 were used.

Techniques: Control, Staining

Journal: JCI insight

Article Title: Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1.

doi: 10.1172/jci.insight.132796

Figure Lengend Snippet: Figure 2. Direct intramyocardial injection of free CHIR99021 and/or FGF1: assessment of cardiac function and infarct size in a mouse model of MI. Mice were treated with direct intramyocardial injections of free CHIR, FGF1, and CHIR + FGF1 while MI-only mice served as controls. (A) Echocardiographic assessment of LV function. EF (B) and FS (C) were determined before MI (pre-S) and after 28 days (post-S). On day 28 after MI, no difference was identified among different treatment groups (means ± SEM, 5 animals per group). (D) Fibrosis, as assessed by Sirius Red/Fast Green histochemical stain- ing (scale bar: 1 mm). At day 28, fibrosis, expressed as the ratio of the scar area to the total surface area of the left ventricle, did not differ between groups (E). There were 5 animals per group. Statistical analysis: 2-way ANOVA (B and C) and 1-way ANOVA (E) with Dunn’s multiple comparisons test. *P < 0.01 vs. sham (B and C, post-S).

Article Snippet: A double-emulsion (water/oil/water phase) technique for recombinant human FGF1 acidic protein (rhFGF1, aa 16-155 (Novus Biologicals, Inc.) and a single-emulsion (oil/water phase) technique for CHIR99021 were used.

Techniques: Injection, Staining

Journal: JCI insight

Article Title: Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1.

doi: 10.1172/jci.insight.132796

Figure Lengend Snippet: Figure 3. Detection and quantification of apoptosis by TUNEL assay in the mouse model of MI. (A) Seventy-two hours posttreatment, the CHIR + FGF1- NP treatment group showed significantly fewer numbers of TUNEL+ cells in the border zone compared with the other treatment groups (scale bar: 50 μm) (B). Cells were also counterstained for nuclei (blue, DAPI). Data are given as means ± SEM. There were 5 animals per group. Statistical analysis: 1-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. sham; †P < 0.01 vs. MI; ‡P < 0.01 vs. empty NP; §P < 0.01 vs. CHIR-NPs; ||P < 0.01 vs. FGF1-NPs.

Article Snippet: A double-emulsion (water/oil/water phase) technique for recombinant human FGF1 acidic protein (rhFGF1, aa 16-155 (Novus Biologicals, Inc.) and a single-emulsion (oil/water phase) technique for CHIR99021 were used.

Techniques: TUNEL Assay

Journal: JCI insight

Article Title: Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1.

doi: 10.1172/jci.insight.132796

Figure Lengend Snippet: Figure 4. Evaluation of NPs mediated neo-angiogenesis in the mouse model of MI. (A) Expression of the endothelium-specific lectin, isolectin B4 (red, IB4) and smooth muscle protein, SM22-α (green, SM22α), in the periinfarct border zone sections 28 days after MI (scale bar: 50 μm). (B and C) Graphical expression of angiogenesis data. The CHIR + FGF1-NP–treated group showed significantly elevated vascular densities compared with other NP treatment groups and the untreated MI group, but no significant difference in vascular densities was observed between the CHIR + FGF1-NP treat- ment group and the sham-operated controls. Data are given as means ± SEM. There were 10–12 animals per group. Statistical analysis: 1-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. sham; †P < 0.01 vs. MI; ‡P < 0.01 vs. empty NPs; §P < 0.01 vs. CHIR-NPs; ||P < 0.01 vs. FGF1-NPs. (D–G) Cell cycle activity of endothelial cells assessed by a dual staining for Ki-67 (green) and IB4 (red; D and F) and phospho–histone H3 (PH3) (green) and IB4 (red; E and G). Scale bar: 20 μm. Data are given as means ± SEM. There were 10–12 animals per group. Statistical analysis: 1-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. sham; †P < 0.01 vs. MI; ‡P < 0.01 vs. empty NPs; §P < 0.01 vs. CHIR-NPs; ||P < 0.01 vs. FGF1-NPs.

Article Snippet: A double-emulsion (water/oil/water phase) technique for recombinant human FGF1 acidic protein (rhFGF1, aa 16-155 (Novus Biologicals, Inc.) and a single-emulsion (oil/water phase) technique for CHIR99021 were used.

Techniques: Expressing, Activity Assay, Staining

Journal: JCI insight

Article Title: Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1.

doi: 10.1172/jci.insight.132796

Figure Lengend Snippet: Figure 5. Assessment of LV morphology and function in a pig model of IR injury. Cardiac MRI recordings in the experimental groups at end-diastole (ED) and end-systole (ES). At day 28, the CHIR + FGF1-NP-treated group revealed significant reduction in infarct size compared with the untreated IR group (B). On the contrary, EF (C), CO (D), and SV (E) were significantly greater in CHIR + FGF1-NP–treated groups than in untreated IR groups while LV end-diastolic volume and LV end-systolic volume were significantly lower (F and G). Data are given as means ± SEM. There were 4 animals per group. Statistical analysis: 2-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. pre-IR; †P < 0.05 vs. week 1; ‡P < 0.05 vs. IR. Macroscopic areas of infarction/fibrosis/scar (H) at after-IR day 28 in serial transverse sections of fresh (scale bar: 1 cm) representative micrographs of Sirius Red/Fast Green histochemical staining, revealing areas of infarcted (red, nonviable) and noninfarcted (green, viable) zones (I) (scale bar: 1 mm) and quantification of left anterior wall thickness (J). Data are given as means ± SEM. There were 4 animals per group. Statistical analysis: 1-way ANOVA with Dunn’s multiple comparisons test. *P < 0.01 vs. control; †P < 0.05 vs. IR.

Article Snippet: A double-emulsion (water/oil/water phase) technique for recombinant human FGF1 acidic protein (rhFGF1, aa 16-155 (Novus Biologicals, Inc.) and a single-emulsion (oil/water phase) technique for CHIR99021 were used.

Techniques: Staining, Control