Journal: Cardiovascular Toxicology

Article Title: Liraglutide Modulates Zinc Release and Improves Mitochondrial Function in Insulin-Resistant Senescent Cardiomyocytes

doi: 10.1007/s12012-026-10095-x

Figure Lengend Snippet: Expression levels of integrated stress markers ATF4, ATF6, and CHOP in insulin-resistant aged cells. A Relative mRNA expression levels of ER stress markers ATF4, ATF6, and CHOP in insulin resistant senescent AC16 cells were treated with Liraglutide and CX-4945 for 30 min. Data are normalized to B-Actin mRNA levels and expressed as fold changes relative to the Con group. Results are given as mean ± SEM, n (Con) :4, n (PA) : 4, n (P+D-Gal) :4, n (PD+Lirag) :4 and n (PDL+CX-4945) :4 number of replicates. B Representative western blot images (top) and quantitative summary data (bottom) showing ATF4, ATF6, and DDIT3 (CHOP) protein levels in the same experimental groups ( n = 4). Results are presented as mean ± SEM. n (Con) :4, n (PA) : 4, n (P+D-Gal) :4, n (PD+Lirag) :4 and n (PDL+CX-4945) :4 number of replicates. Statistical comparisons were performed using the Kruskal-Wallis test. * p < 0.05 vs. Con, # p < 0.05 vs. P + D-Gal, $ p < 0.05 vs. PD + Lirag.

Article Snippet: Proteins were transferred (Bio-Rad – Trans-Blot Turbo) to PVDF membranes and blocked with 3% BSA for 90 min. Membranes were probed overnight with monoclonal (mono) or polyclonal (poly) primary antibodies diluted in 3% BSA for P62 (814801; 1:500; BioLegend), Calnexin (sc-23954; 1:500; Santa Cruz), LC3 (848802, 1:500, BioLegend), ATF4 (STJ11101481; 1:500; BioLegend), ATF6 (STJ114444; 1:500; St John’s Laboratory), GRP78 (sc-13968; 1:500; Santa Cruz), B-actin (664802; 1:500; BioLegend), and GAPDH (649202, 1:500; BioLegend) as loading controls.

Techniques: Expressing, Western Blot

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Relationship between cyclooxygenase-2 and unfolded protein response pathways. Expression of unfolded protein response (UPR) pathways and COX-2 in hepatocellular carcinoma (HCC) (streptavidin-peroxidase × 400). A: Expression of COX-2 in HCC; B: Expression of ATF-6 in HCC; C: Expression of IRE-1 in HCC; D: Expression of PERK in HCC. COX-2: Cyclooxygenase-2.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Expressing

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Selection of the proper melatonin concentration. The effects of melatonin (MT) on the three unfolded protein response (UPR) pathways of HepG2 cells induced by tunicamycin (TM). HepG2 cells were exposed to different concentrations of melatonin (10 -9 , 10 -7 , 10 -5 and 10 -3 mol/L) for 24 h. A: Equal protein amounts of cell lysates were subjected to western blot assay using anti-ATF-6, anti-IRE-1 and anti-PERK. β-actin in the same HepG2 cells extract was used as an internal reference; B: Optical density reading values of specific proteins are represented as fold differences relative to the loading control protein, β-actin. a P < 0.01 vs negative control (NC); b P < 0.05 vs positive control (TM); c P < 0.01 vs positive control (TM). ATF-6: Activating transcription factor 6; IRE: Inositol-requiring enzyme; PERK: Protein kinase RNA-like endoplasmic reticulum kinase.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Selection, Concentration Assay, Western Blot, Negative Control, Positive Control

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Selection of the most effective sequences of each unfolded protein response pathways siRNA (A-C). All of the candidate sequences of the three unfolded protein response (UPR) pathways were examined by RNA interference, and the expression of UPR proteins was evaluated by western blotting to select the most effective one to interfere with the UPR pathway. ATF-6: Activating transcription factor 6; NC: Negative control; IRE: Inositol-requiring enzyme; PERK: Protein kinase RNA-like endoplasmic reticulum kinases.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Selection, Expressing, Western Blot, Negative Control

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Relationship between activating transcription factor 6 and cyclooxygenase-2 under endoplasmic reticulum stress. The mRNA of the unfolded protein response (UPR) pathways was interfered by ATF-6, IRE-1 and PERK siRNA after pretreatment by tunicamycin (TM) for 8 h. A: Equal protein amounts of cell lysates were subjected to western blot assay using anti-COX-2. β-actin in the same HepG2 cell extract was used as an internal reference. B: Optical density reading values of specific proteins are represented as fold-differences relative to the loading control protein, β-actin. a P < 0.01 vs the negative control. RNA was harvested and gene expression examined by qRT-PCR in the same condition above. C: The qRT-PCR fold-changes were normalized using the expression of housekeeping gene (GAPDH) and vs those obtained from untreated HepG2 cells. a P < 0.01 vs the negative control; c P < 0.01 vs the untreated HepG2 cells. ATF-6: Activating transcription factor 6; NC: Negative control; COX-2: Cyclooxygenase-2; IRE: Inositol-requiring enzyme; PERK: Protein kinase RNA-like endoplasmic reticulum kinase.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Western Blot, Negative Control, Expressing, Quantitative RT-PCR

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Effects of activating transcription factor 6 siRNA on cell apoptosis in HepG2 cells under endoplasmic reticulum stress. A: HepG2 cells were transfected with ATF-6 siRNA for 24 h after pretreatment by tunicamycin (TM) for 8 h. Apoptotic cells were determined by FACS, and the data are expressed as the mean ± SD. A1, Untreated HepG2 cells; A2, HepG2 cells treated by TM only; A3, HepG2 cells treated by combination of TM and ATF-6 siRNA negative control; A4, HepG2 cells treated by ATF-6 siRNA and melatonin; B: Data are presented as the mean ± SD for the independent experiments ( a P < 0.05 vs untreated HepG2 cells; b P < 0.01 vs untreated HepG2 cells; c P < 0.01 vs HepG2 cells treated with TM and ATF-6 siRNA negative control); C: Cell morphology and percentage of apoptotic cells was examined by TUNEL staining. C1, Untreated HepG2 cells; C2, HepG2 cells treated by TM only; C3, HepG2 cells treated by combination of TM and ATF-6 siRNA negative control; C4, HepG2 cells treated by ATF-6 siRNA and melatonin; D: Data are presented as the mean ± SD for the independent experiments ( b P < 0.01 vs untreated HepG2 cells; c P < 0.01 vs HepG2 cells treated with TM and ATF-6 siRNA negative control). ATF-6: Activating transcription factor 6; NC: Negative control.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Transfection, Negative Control, TUNEL Assay, Staining

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Comparison between melatonin and activating transcription factor 6 siRNA transfection on cell apoptosis in HepG2 cells under endoplasmic reticulum stress. A: HepG2 cells were exposed to melatonin (10 -5 mmol/L) for 24 h after pretreatment by tunicamycin (TM) for 8 h or were transfected with ATF-6 siRNA for 6 h after pretreatment by tunicamycin TM for 8 h. Apoptotic cells were determined by FACS, and the data are expressed as the mean ± SD. A1, Untreated HepG2 cells; A2, HepG2 cells treated by TM only; A3, HepG2 cells treated by combination of TM and melatonin (10 -5 mmol/L); A4, HepG2 cells treated by ATF-6 siRNA and melatonin; B: Data are presented as the mean ± SD for the independent experiments ( a P < 0.05 vs untreated HepG2 cells; b P < 0.01 vs untreated HepG2 cells; c P < 0.01 HepG2 cells treated by TM and melatonin vs HepG2 cells treated with TM and ATF-6 siRNA negative control); C: Cell morphology and percentage of apoptotic cells were examined by TUNEL staining. C1, Untreated HepG2 cells; C2, HepG2 cells treated by TM only; C3, HepG2 cells treated by combination of TM and melatonin (10 -5 mmol/L); C4, HepG2 cells treated by ATF-6 siRNA and melatonin; D: Data are presented as the mean ± SD for the independent experiments ( a P < 0.05 vs untreated HepG2 cells; b P < 0.01 vs untreated HepG2 cells; c P < 0.01 HepG2 cells treated by TM and melatonin vs HepG2 cells treated with TM and ATF-6 siRNA negative control). ATF-6: Activating transcription factor 6; MT: Melatonin.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Transfection, Negative Control, TUNEL Assay, Staining

Journal: World Journal of Gastroenterology

Article Title: Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation

doi: 10.3748/wjg.v23.i6.986

Figure Lengend Snippet: Pathway by which melatonin sensitized HepG2 to endoplasmic reticulum stress-induced apoptosis. A: a, Untreated HepG2 cells; b, HepG2 cells treated by tunicamycin (TM) only; c, HepG2 cells treated by TM, melatonin (10 -5 mmol/L); d: HepG2 cells treated by TM and ATF-6 siRNA. Equal protein amounts of cell lysates were subjected to western blot assay using anti-COX-2, anti-CHOP, anti-Bcl-2, and anti-Bax. β-actin in the same HepG2 cell extract was used as an internal reference; B: Optical density reading values of specific proteins are represented as fold-differences relative to the loading control protein, β-actin. c P < 0.01, CHOP vs the Bcl-2 and Bax expression, b P < 0.01, vs the untreated HepG2 cells. ATF-6: Activating transcription factor 6; COX-2: Cyclooxygenase-2; CHOP: CCAAT-enhancer-binding protein homologous protein; MT: Melatonin.

Article Snippet: Anti-ATF-6 (BS6476), anti-PERK (BS2156) and anti-Bcl-2 (BS3711) were obtained from Bioworld Technology Inc. (St Louis Park, MN, United States).

Techniques: Western Blot, Expressing, Binding Assay