Journal: Pharmaceutics

Article Title: Insights into the Structural Patterns in Human Glioblastoma Cell Line SF268 Activity and ADMET Prediction of Curcumin Derivatives

doi: 10.3390/pharmaceutics17080968

Figure Lengend Snippet: Immunofluorescence staining of microtubules in SF268 cells. Cells were treated for 12 h with 0.5% DMSO as an untreated control ( A ), 10 μM staurosporine ( B ) as an apoptosis control, and the following curcumin derivatives (10 μg/mL): curcumin ( C ), compound 2 ( D ), compound 6 ( E ), and compound 11 ( F ). The cells were then stained with a β-tubulin antibody (yellow) and Hoechst dye (blue) to visualize the nucleus.

Article Snippet: A Tubulin Polymerization Assay Kit (Porcine tubulin and Fluorescence-based; Cat. # BK011P) (Cytoskeleton Inc., Denver, CO, USA) was used.

Techniques: Immunofluorescence, Staining, Control

Journal: Pharmaceutics

Article Title: Insights into the Structural Patterns in Human Glioblastoma Cell Line SF268 Activity and ADMET Prediction of Curcumin Derivatives

doi: 10.3390/pharmaceutics17080968

Figure Lengend Snippet: ( A ) Tubulin polymerization of compounds 1 , 2 , 3 , 4 , 6 , and 11 . Tubulin polymerization was monitored by the increase in fluorescence at 360 nm (excitation) and 420 nm (emission) for 1 h at 37 °C. Paclitaxel and calcium chloride were used as positive controls, whereas 0.1% DMSO was used as a negative control. ( B ) AUC for the tested compounds and positive and negative controls. **** p ≤ 0.0001.

Article Snippet: A Tubulin Polymerization Assay Kit (Porcine tubulin and Fluorescence-based; Cat. # BK011P) (Cytoskeleton Inc., Denver, CO, USA) was used.

Techniques: Fluorescence, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Identified Three Interferon Induced Proteins as Novel Biomarkers of Human Ischemic Cardiomyopathy

doi: 10.3390/ijms222313116

Figure Lengend Snippet: Cardiac function evaluation in rats after continuous administration Metoprolol. ( A ) The echocardiographic analysis of different groups, including Sham, myocardial infarction (Lad) and Met group. ( B ) The average of EF and FS in the Met group were significantly higher than Lad group. Sham group: n = 10; Lad group n = 8; Met group: n = 10. ( C ) The RT-qPCR to test gene expression in left ventricle (Sham group: n = 5; Lad group: n = 6; Met group: n = 8). ( D ) The proteins in MI rat serum were detected by ELISA (Sham group: n = 10; Lad group: n = 8; Met group: n = 10). ( E ) The ratios of proteins. The proportion of IFIT2/IFIT3 in sham group, Lad group and Met group were 1.066 ± 0.08, 1.076 ± 0.05 and 1.104 ± 0.05, respectively. The ratio of BCL2L1/IFIT3 was significantly increased in LAD group (2.399 ± 0.07), compared with Sham group (1.715 ± 0.07) and MET group (1.859 ± 0.05). The proportion of IFI44L/IFIT2 was similar with BCL2L1/IFIT3 (Sham group, 1.07 ± 0.04; LAD group, 1.27 ± 0.06; MET group, 1.055 ± 0.05). Sham group: n = 10; Lad group: n = 11; Met group: n = 12. ( F ) Cardiomyocyte’s apoptosis in infract zone. DAPI channel showed blue fluorescence and TUNEL-positive cardiomyocytes indicated green fluorescence and the merged channel was DAPI and TUNEL. LAD group, 55.50 ± 0.72; MET group, 33.15 ± 4.45. p = 0.0077, n = 3. All results were expressed as the means ± SEM and a dependent t -test would be used. * Represents p < 0.05; ** represents p < 0.01; *** represents p < 0.001; **** represents p < 0.0001.

Article Snippet: After being washed three times, the sections were incubated with TUNEL reagents (One step TUNEL apoptosis assay kit, green fluorescence, Cat NO.1088, Beyotime) for one hour at 37 °C.

Techniques: Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay, Fluorescence, TUNEL Assay

Journal: Cell Cycle

Article Title: Inflammatory stimuli promote growth and invasion of pancreatic cancer cells through NF-κB pathway dependent repression of PP2Ac

doi: 10.1080/15384101.2015.1127468

Figure Lengend Snippet: Inflammatory stimuli promoted growth and invasion of pancreatic cancer cells in vitro. (A) MTT assay of viability of cells incubated with LPS (0, 10, 100, 1000 ng/ml) for 24, 48, and 72 h. (B) Cell invasion assay following 24-h exposure to 10 ng/ml LPS. Cells that had migrated to the lower membranes were photographed under ×400 magnification. **P < 0.01 as compared to control. (C) Quality control of MCM. After 7-day MCSF treatment, IL-8 and TNF-α levels at 24, 48, 72, 96, and 120 h were measured using Milliplex assay. (D) MTT assay of viability of cells incubated with MCM for 24, 48, 72, 96, and 120 h. *P < 0.05 as compared with control. (E) Cell invasion assay following 24-h exposure to MCM. Cells that had migrated to the lower membranes were photographed under ×400 magnification. **P < 0.01 as compared to control.

Article Snippet: Multiplex biometric immunoassay kits containing fluorescent dyed microbeads (Milliplex NF-κB Signaling Magnetic Bead kit; Cat. No. 48-630MAG, and Human Cytokine MAGNETIC Kit; Cat. No. HCYTOMAG-60K, Millipore Corp, St Charles, MO) were used for measuring phosphorylated IKKα/β (Ser176/Ser180), phosphorylated IκBα (Ser32), c-Myc, IL-8, and TNF-α.

Techniques: In Vitro, MTT Assay, Incubation, Invasion Assay, Control

Journal: Cell Cycle

Article Title: Inflammatory stimuli promote growth and invasion of pancreatic cancer cells through NF-κB pathway dependent repression of PP2Ac

doi: 10.1080/15384101.2015.1127468

Figure Lengend Snippet: Inflammatory stimuli induced NF-κB pathway activation in pancreatic cancer cells. (A, B) Milliplex assay evaluation of IKK phosphorylation at Ser176/Ser180 in SW1990 and CFPAC-1 cells treated with 10 ng/ml LPS (A) or MCM (B) for 12 h. (C) DN-IKKα (S176/180A) overexpression repressed LPS-induced IKK phosphorylation in SW1990 (top) and CFPAC-1 (bottom) cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05 indicates significant differences between fold induction. (D) DN-IKKα (S176/180A) overexpression repressed MCM-induced IKK phosphorylation in SW1990 cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05 indicates significant differences between fold induction. (E, F) Milliplex assay evaluation of IκBα phosphorylation at Ser32 after 12-h treatment with 10 ng/ml LPS (E) or MCM (F). (G) DN-IKKα (S176/180A) or DN-IκBα (S32/36A) overexpression repressed LPS-induced IκBα phosphorylation in SW1990 (top) and CFPAC-1 (bottom) cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05, and&P < 0.01 indicate significant differences between fold induction. (H) DN-IKKα (S176/180A) or DN-IκBα (S32/36A) overexpression repressed MCM-induced IκBα phosphorylation in SW1990 cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05, and&P < 0.01 indicate significant differences between fold induction. (I) Western blot of p65 expression in cells treated with 10 ng/ml LPS for 12 h, followed by total and nuclear protein extraction. β-actin and histone H1 were the internal controls for the total and nuclear extracts, respectively. (J, K) Luciferase reporter gene assays of cells transiently transfected with pNF-κB-luc before 36-h treatment with 10 ng/ml LPS (J) or MCM (K). **P < 0.01 as compared with control. (L, M) Milliplex assay of c-Myc expression in cells treated with LPS (L) or MCM (M) for 12 h. **P < 0.01 as compared with control.

Article Snippet: Multiplex biometric immunoassay kits containing fluorescent dyed microbeads (Milliplex NF-κB Signaling Magnetic Bead kit; Cat. No. 48-630MAG, and Human Cytokine MAGNETIC Kit; Cat. No. HCYTOMAG-60K, Millipore Corp, St Charles, MO) were used for measuring phosphorylated IKKα/β (Ser176/Ser180), phosphorylated IκBα (Ser32), c-Myc, IL-8, and TNF-α.

Techniques: Activation Assay, Phospho-proteomics, Over Expression, Control, Western Blot, Expressing, Protein Extraction, Luciferase, Transfection