Journal: Experimental and molecular pathology

Article Title: Hepatic alcohol dehydrogenase deficiency induces pancreatic injury in chronic ethanol feeding model of deer mice

doi: 10.1016/j.yexmp.2018.01.002

Figure Lengend Snippet: Average body weights of ADH− and ADH+ deer mice during four months of ethanol feeding. P ≤ 0.05 for ethanol-fed ADH− vs. ethanol-fed ADH+ deer micea, ethanol-fed ADH− vs. pair-fed control ADH− deer miceb and ethanol-fed ADH+ vs. pair-fed control ADH+ deer micec. * - initial body weight of the mice before feeding 1g% ethanol in the liquid diet. The ethanol concentration was gradually increased to 3.5g% in the liquid diet in two weeks period and maintained for remaining 120 days (n = 10 animals/group).

Article Snippet: Polyclonal antibodies raised against a synthetic peptide corresponding to the sequence of human glucose regulated protein (GRP)78 (Cell Signaling, Danvers, MA; Cat# 3183; 1:1000 dilution; MW, 78 kDa), inositol-requiring enzyme (IRE)1α (Novus Biologicals, Littleton, CO; Cat# NB100-2323; 1:1000 dilution; MW, 110 kDa), activating transcription factor (ATF) 6 (Novus Biologicals; Cat# nbp1-40256; 5μg/ml dilution; MW, 90 kDa: full length & 50kDa: active/cleaved), phosphorylated eukaryotic initiation factor (p-eIF)2α (Cell Signaling, Cat# 9721; 1:1000 dilution; MW, 38 kDa), X-box binding protein (XBP)1s (Abcam, Cambridge, MA; Cat# ab37152; 2μg/ml dilution; MW, 29 kDa [isoform 1] & 42 kDa [isoform 2]), proapoptotic protein CCAAT/-enhancer-binding protein homologous protein (CHOP) (Cell Signaling, Cat# 5554; 1:1000 dilution; MW, 27 kDa), phosphorylated protein kinase R-like endoplasmic reticulum kinase (PERK) (Santa Cruz, Dallas, TX; Cat# sc-32577; 1:1000 dilution; MW, 125 kDa), 4-hydroxynonenal (4-HNE) (Abcam, Cat# ab46545; 1:200 dilution), and anti-rabbit IgG and HRP-linked secondary antibody (Cell Signaling, Cat# 7074; 1:1000 dilution) were procured.

Techniques: Control, Concentration Assay

Journal: Nutrients

Article Title: Role of Endoplasmic Reticulum and Oxidative Stress Parameters in the Pathophysiology of Disease-Related Malnutrition in Leukocytes of an Outpatient Population

doi: 10.3390/nu11081838

Figure Lengend Snippet: Leukocyte expression of ER stress proteins in NN and DRM subjects. ( A ) CHOP protein expression relative to actin protein expression, in NN vs DRM patients. ( B ) ATF6 protein expression relative to actin protein expression in NN vs DRM patients. ( C ) P-Eif2α protein expression relative to actin protein expression in NN vs DRM patients. ( D ) GRP78 protein expression relative to actin protein expression in NN vs DRM patients. * p < 0.05 vs. NN group using an unpaired t -test NN: Normonutrition; DRM: Disease-related malnutrition; ER: endoplasmic reticulum

Article Snippet: The primary antibodies (anti-GRP78 rabbit polyclonal antibody (Abcam, Cambridge, UK), anti-ATF6 mouse monoclonal antibody (which detects the cleaved/active form of ATF6; Thermo Scientific, Rockford, IL, USA), anti-CHOP mouse monoclonal antibody (Thermo Scientific, Rockford, IL, USA), anti-eIF2α-pS52 (Life Technologies, Barcelona, Spain), and anti-actin rabbit polyclonal antibody (Sigma Aldrich, St. Louis, MO, USA) were incubated overnight at 4 °C under gentle shaking.

Techniques: Expressing

Journal: Journal of Diabetes Research

Article Title: 3 β -Hydroxysteroid- Δ 24 Reductase (DHCR24) Protects Pancreatic β Cells from Endoplasmic Reticulum Stress-Induced Apoptosis by Scavenging Excessive Intracellular Reactive Oxygen Species

doi: 10.1155/2020/3426902

Figure Lengend Snippet: UPR-related signaling pathways were detected after TM treatment by Western blot analysis. (a) Representative Western blots show the expression of c-myc, Bip, p-p38, ATF6, and cleaved ATF6 in MIN6 cells after TM treatment. The levels of these proteins were normalized to actin levels. (b, c) The levels of Bip, p-p38, ATF6, and cleaved ATF6 were calculated as percentage of the largest level in the negative control (Ad-LacZ) group. Mean ± S.D. ( n = 3); ∗ p < 0.05 vs. Ad-LacZ.

Article Snippet: We used the following primary antibodies: rabbit anti-actin (Sigma-Aldrich, St. Louis, MO), mouse anti-Bip/GRP78 (BD Biosciences, Bedford, MA), mouse anti-myc (Santa Cruz, CA, USA), mouse anti-ATF6/cleaved ATF6, and rabbit anti-phospho-p38 MAPK (T180/Y182) (Cell Signaling, Beverly, MA).

Techniques: Protein-Protein interactions, Western Blot, Expressing, Negative Control

Journal: Frontiers in Immunology

Article Title: Hyperglycemia Aggravates Hepatic Ischemia and Reperfusion Injury by Inhibiting Liver-Resident Macrophage M2 Polarization via C/EBP Homologous Protein-Mediated Endoplasmic Reticulum Stress

doi: 10.3389/fimmu.2017.01299

Figure Lengend Snippet: Hyperglycemia enhances endoplasmic reticulum stress in livers post-IR. Diabetic [streptozotocin (STZ)] and control (CON) mice were prepared and liver partial warm ischemia and reperfusion (IR) or sham procedure was performed. Six hours post-reperfusion, liver tissues were collected, and c-ATF6, ATF4, C/EBP homologous protein (CHOP), s-XBP1, and β-actin protein levels were analyzed by Western blotting. Representative of three experiments (A) . Relative density ratios of target proteins in different groups to the control group (CON-Sham) were calculated [ (B) , n = 3/group] (* p < 0.05).

Article Snippet: Primary antibodies against cleaved-ATF6 (c-ATF6, Novus, Littleton, CO, USA), ATF4 (Proteintech Group, Chicago, IL, USA), CHOP (Cell Signaling Technology, MA, USA), spliced XBP1 (s-XBP1, Abcam, Cambridge, MA, USA), and β-actin (Cell Signaling Technology, MA, USA) were used and incubated overnight at 4°C.

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Hyperglycemia Aggravates Hepatic Ischemia and Reperfusion Injury by Inhibiting Liver-Resident Macrophage M2 Polarization via C/EBP Homologous Protein-Mediated Endoplasmic Reticulum Stress

doi: 10.3389/fimmu.2017.01299

Figure Lengend Snippet: C/EBP homologous protein (CHOP) mediates hyperglycemic Kupffer cell (KC) pro-inflammatory activation in vitro . Diabetic [streptozotocin (STZ)] and control (CON) mice were prepared and liver partial warm ischemia and reperfusion (IR) or a sham procedure was performed. After 6 h of reperfusion, KCs were isolated and the intracellular levels of c-ATF6, ATF4, CHOP, s-XBP1, and β-actin protein were analyzed by Western blotting. Representative of three experiments (A) . Relative density ratios of target proteins in different groups to the control group (CON-Sham) were calculated [ (B) , n = 3/group]. Both CON and STZ mice were pretreated with CHOP siRNA (CHOP-siRNA) or its scramble control siRNA (SCR-siRNA) in vivo prior to IR using mannose-conjugated polymers as described in Section “ .” Liver IR was performed. Six hours post-reperfusion, KCs were isolated and the intracellular levels of c-ATF6, ATF4, CHOP, s-XBP1, and β-actin protein were analyzed by Western blotting. Representative of three experiments (C) . Relative density ratios of target proteins in different groups to the control group (CON–SCR-siRNA) were calculated [ (D) , n = 3/group]. Isolated KCs from IR-stressed livers of different groups were cultured for 6 h, and TNF-α, IL-6, and IL-10 protein levels in the culture supernatant were measured by ELISA [ (E) , n = 6/group] (* p < 0.05).

Article Snippet: Primary antibodies against cleaved-ATF6 (c-ATF6, Novus, Littleton, CO, USA), ATF4 (Proteintech Group, Chicago, IL, USA), CHOP (Cell Signaling Technology, MA, USA), spliced XBP1 (s-XBP1, Abcam, Cambridge, MA, USA), and β-actin (Cell Signaling Technology, MA, USA) were used and incubated overnight at 4°C.

Techniques: Activation Assay, In Vitro, Isolation, Western Blot, In Vivo, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Argininosuccinate synthase 1 suppresses tumor progression through activation of PERK/eIF2α/ATF4/CHOP axis in hepatocellular carcinoma

doi: 10.1186/s13046-021-01912-y

Figure Lengend Snippet: ASS1 controls cell fate through upregulation of CHOP in HCC a Expression of ER stress-related proteins; PERK, ATF6, IRE1a, CHOP, ATF3 XBP1s and XBP1u in pCMV3 or ASS1-flag-transfected Huh7 and SNU475 cells. b Expression of apoptosis-related proteins; PARP and caspase 3 active forms and ER stress related proteins; CHOP and GRP78 in Huh7 and SNU475 cells after treatment with TG. Cell viability analysis of TG treatment in c pCMV3 or ASS1-flag transfected SNU475 cells and d siRNA against ASS1 transfected Hep3B cells. e The colony formation assay and f The expression of apoptosis related proteins in Huh7 and SNU475 cells transfected with either non-specific control siRNA (siCont.) or CHOP siRNA (siCHOP) in ASS1 overexpressed Huh7 and SNU475 cells. g Expression of ASS1 and CHOP in ASS1 overexpressed Huh7 cell transfected with non-specific siRNA (siCont.) or ASS1 siRNA (siASS1) by western blot assay. h The correlation expression levels between ASS1 and CHOP in tumor tissues from HCC patients. Immunofluorescence (IF) staining of ASS1 and CHOP in i Tumor tissues from patients with liver cancer and j Huh7 cells. k Co-immunoprecipitation (IP) assay of Huh7 cells transfected with ASS1-Flag. The samples were analyzed by immunoblotting with an anti-PERK, ATF4, and CHOP. * P < 0.05 compared to siControl group

Article Snippet: Primary antibodies for hybridization included those for ATF6 (65880S), IRE1α (3294S), ATF3 (33593S), GRP78 (3117S), cleaved caspase 3 (7237S), PARP (5625S), Snail (3879S) from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Transfection, Colony Assay, Control, Western Blot, Immunofluorescence, Staining, Immunoprecipitation

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: Gene primer sequences.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: Effects of GC on CP-induced ER stress. (A) Effect of GC on the morphological changes of ER in liver cells induced by CP. Red arrows show the morphology of the endoplasmic reticulum and ribosomes. (B, C) Effect of GC on CP-induced ER stress-related indicators. GRP78, ATF6, and p-IRE1α protein expression were detected by WB analysis. GRP78 and ATF6 gene expression were detected by qRT-PCR analysis. All data are presented as the mean ± SD. ## p < 0.01 compared with control group; * p < 0.05, ** p < 0.01 compared with CP group.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Control

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: (A-C) MTT assay for determining cell viability. (D-E) GC reduced the expression of ER stress-related indicators. Expression levels of GRP78, ATF6, and p-IRE1α protein were tested by WB analysis. Expression levels of GRP78 and ATF6 mRNA were tested by qRT-PCR analysis.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques: MTT Assay, Expressing, Quantitative RT-PCR