Journal: Antioxidants

Article Title: Identification of Dietary Phytochemicals Capable of Enhancing the Autophagy Flux in HeLa and Caco-2 Human Cell Lines

doi: 10.3390/antiox9121193

Figure Lengend Snippet: Establishment of an assay system for detecting cellular autophagy flux. HeLa cells stably expressing green fluorescent protein (GFP)-microtubule-associated protein light chain 3 (LC3)-red fluorescent protein (RFP)-LC3ΔG were treated with Torin1 (250 nM) or bafilomycin A1 (BAF; 100 nM) for 12 h ( A , B ). Probe expression was detected by fluoroimaging ( A ) and western blotting using antibodies against GFP or RFP ( B ); β-Actin was used as an internal control. Using an Operetta high-content imaging system, co-cultured transgenic and wild-type HeLa cells were distinguished by digital phase-contrast and RFP fluorescence (yellow, wild-type; red, transgenic); the net fluorescence derived from the probe was calculated to obtain the GFP/RFP ratio ( C ).

Article Snippet: The mouse monoclonal antibody against SQSTM1 (D3; sc-28359) and the mouse polyclonal antibody against β-actin (C4; sc-4778) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Stable Transfection, Expressing, Western Blot, Control, Imaging, Cell Culture, Transgenic Assay, Fluorescence, Derivative Assay

Journal: Antioxidants

Article Title: Identification of Dietary Phytochemicals Capable of Enhancing the Autophagy Flux in HeLa and Caco-2 Human Cell Lines

doi: 10.3390/antiox9121193

Figure Lengend Snippet: Effects of active phytochemicals on p62 protein expressions in Caco-2 cells. Caco-2 cells were treated with Torin1 (1 μM) or each flavonoid (20 μM) for 4 h. To determine the autophagy flux, intracellular fluorescence intensities were measured using a Cellometer ® Vision instrument to obtain GFP/RFP ratios ( A ). p62 protein level was assessed by western blotting. β-actin was used as an internal control ( B , C ). Data represent the mean ± SEM of triplicate measurements. Different letters indicate significant differences as assessed by the Tukey–Kramer test ( p < 0.05) ( C ). Zer, zerumbone; IsoR, isorhamnetin; Chr, chrysoeriol; THC, 2,2′,4′-trihydroxychalcone.

Article Snippet: The mouse monoclonal antibody against SQSTM1 (D3; sc-28359) and the mouse polyclonal antibody against β-actin (C4; sc-4778) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Fluorescence, Western Blot, Control

Journal: Antioxidants

Article Title: Identification of Dietary Phytochemicals Capable of Enhancing the Autophagy Flux in HeLa and Caco-2 Human Cell Lines

doi: 10.3390/antiox9121193

Figure Lengend Snippet: mTOR signaling contribution to the autophagy-promoting effects of phytochemicals. Transgenic Caco-2 cells were treated with Torin1 (1 μM) and/or each flavonoid (20 μM) for 4 h. To determine the autophagy flux, intracellular fluorescence intensities were measured using a Cellometer ® Vision instrument to obtain GFP/RFP ratios ( A ). mTORC1 activity was assessed by determining the level of phospho-p70 S6K and 4EBP1 band shifts. β-Actin was used as an internal control ( B , C ). Data represent the mean ± SEM of triplicate measurements. Statistical significance was assessed by unpaired t -test (* p < 0.05) ( A ). The different letters indicate significant differences as determined using the Tukey–Kramer test ( p < 0.05) ( C ). Zer, zerumbone; IsoR, isorhamnetin; Chr, chrysoeriol; THC, 2,2′,4′-trihydroxychalcone.

Article Snippet: The mouse monoclonal antibody against SQSTM1 (D3; sc-28359) and the mouse polyclonal antibody against β-actin (C4; sc-4778) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Transgenic Assay, Fluorescence, Activity Assay, Control

Journal: Medicine

Article Title: IL12B and IL17 genes polymorphisms associated with differential susceptibility to juvenile idiopathic arthritis and juvenile-onset systemic lupus erythematosus in Chinese children

doi: 10.1097/MD.0000000000034477

Figure Lengend Snippet: Demographic and clinical characteristics of the study participants.

Article Snippet: Serum rheumatoid factor, Complement C3 and Complement C4 were measured by nephelometry (BN II, SIEMENS, Germany).

Techniques: