Journal: Nature Communications

Article Title: Diet-induced RKIP downregulation disrupts PC/PE-ER homeostasis to drive MASLD

doi: 10.1038/s41467-025-65982-8

Figure Lengend Snippet: a Western blotting analysis of RKIP expression in PA-stimulated HepG2 cells with CHX (100 μg/mL), CQ (100 μM) or MG132 (10 μM) treatment for 12 hrs. b Western blotting analysis of RKIP expression in TM (1.2 μM) or Tha (100 nM)-stimulated HepG2 cells with CHX, MG132 or CQ treatment for 6 hrs. c Western blotting analysis of RKIP expression in PA-stimulated HepG2 cells treated with TUDCA (1 mM) or Vehicle (Veh) for the indicated times. d , e Western blotting analysis of RKIP expression in HepG2 ( d ) and HeLa cells ( e ) treated with PA, PA plus CB-5083 (2.5 μM) or Vehicle for 12 hrs. f , g Anti-VCP Co-Immunoprecipitation (Co-IP) of the interaction between VCP and RKIP in HepG2 ( f ) and HeLa cells ( g ) with or without PA stimulation for 12 hrs. h Acyl-biotin exchange (ABE) assays measuring the acylation levels of RKIP-flag in transfected HepG2 cells with or without PA stimulation for 12 hrs. i The acylation levels of RKIP-flag in transfected HepG2 cells with PA (12 hrs), PA plus TUDCA (12 hrs) or Tha (6 hrs) treatment. j The acylation levels of RKIP-flag, RKIP C133A -flag and RKIP C168A -flag in transfected HEK293T cells. k Western blotting analysis of RKIP-flag, RKIP C133A -flag and RKIP C168A -flag in transfected HEK293T cells with or without MG132 treatment for 12 hrs. l Anti-flag Co-IP of the interaction between RKIP-flag, RKIP C133A -flag or RKIP C168A -flag and VCP in transfected HEK293T cells in the presence of MG132. m Anti-GFP Co-IP of the interaction between ZDHHC4 -GFP (left) or ZDHHC6-GFP (right) and RKIP-flag in co-transfected HEK293T cells treated with PA (12 hrs), PA plus TUCDA (12 hrs), Tha (6 hrs) or TM (6 hrs) in the presence of MG132. n Anti-flag Co-IP of the interaction between RKIP-flag and ZDHHC3/4/6 in RKIP-flag-overexpressed HepG2 cells with or without PA stimulation for 12 hrs in the presence of MG132. Numbers below the blot indicate the results of densitometric analysis of RKIP relative to GAPDH ( a-e , k ). Data are representative of at least two independent experiments. Source data are provided as a Source Data file .

Article Snippet: After plating for 24 hrs, 70% confluent cells were treated with or without palmitic acid (PA) (0.5 mM for primary hepatocytes and HepG2 cells, 0.2 mM for HeLa cells, Sigma, P5585), 0.5 mM Oleic acid (OA), 100 μg/mL Cycloheximide (CHX) (MedChemExpress, HY-12320), 100 μM Chloroquine (CQ) (MedChemExpress, HY-17589), 10 μM MG132 (MedChemExpress, HY-12359), 5 μg/mL Actinomycin D (ActD) (MedChemExpress, HY-17559), 1 mM Tauroursodeoxycholic acid (TUDCA) (Selleck, S3654), 1.2 μM Tunicamycin (TM) (Selleck, S7894), 100 nM Thapsigargin (Tha) (Selleck, S7895), 2.5 μM CB-5083 (MedChemExpress, HY-12861) and 100 μM 2-bromopalmitate (2-BP) (Sigma, 21604).

Techniques: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection

Journal: Nature Communications

Article Title: Diet-induced RKIP downregulation disrupts PC/PE-ER homeostasis to drive MASLD

doi: 10.1038/s41467-025-65982-8

Figure Lengend Snippet: a , b Western blotting ( a ) and quantitative analysis ( b ) of ER stress markers in liver of CD- or HFD-fed RKIP f/f and Alb cre RKIP f/f mice (CD groups, n = 3; HFD groups, n = 4). c , d Western blotting analysis of ER stress markers in WT and RKIP knockout (KO) HepG2 ( c ) and HeLa cells ( d ) with PA stimulation for the indicated times. e Representative fluorescence images of LDs (upper), the percentage of LDs (bottom left, n = 3) and the mean number of LDs per cell (bottom right, n = 20) in WT and KO primary hepatocytes treated with PA, TUDCA, PA plus TUDCA or Vehicle for 24 hrs. Scale bars, 40 μm. f RKIP f/f and Alb cre RKIP f/f mice were fed with HFD for 16 weeks and treated with TUDCA (250 mg/kg/day, i.p.) for the last 4 weeks. (Vehicle, n = 3 per group, TUDCA n = 5 per group) Representative H&E staining, Oil red O staining and Masson staining of the liver tissues. Representative 3 liver sections from 3 mice per group. Scale bars, 200 μm. g Quantification for the LDs percentage of liver sections as described in f . h , i The levels of serum ALT ( h ), hepatic total cholesterol (TC) and triglyceride (TG) ( i ) of HFD-fed and TUDCA-treated RKIP f/f and Alb cre RKIP f/f mice as described in f . Data are presented as mean ± SEM ( b, e, g-i ). * p < 0.05, ** p < 0.01, *** p < 0.001. Each dot represents a biological replicate ( b, g-i ) or a technical replicate ( e ). P values were calculated by two-tailed unpaired Student’s t -tests in ( b, e ) and one-way ANOVA in ( g-i ). The samples were derived from the same experiment, pIRE1, RKIP and GAPDH were probed on the same gel, and IRE1, pPERK, PERK, pEIF2α, EIF2α, p-p65, p65, pERK1/2 and ERK1/2 were processed on separate gels in parallel in ( a, c, d ). Numbers below the blot indicate the results of densitometric analysis of the phosphorylated protein relative to the total protein (P/T) and the indicated protein relative to GAPDH (/GAPDH) ( c, d ). Data are representative of at least two independent experiments. Source data are provided as a Source Data file .

Article Snippet: After plating for 24 hrs, 70% confluent cells were treated with or without palmitic acid (PA) (0.5 mM for primary hepatocytes and HepG2 cells, 0.2 mM for HeLa cells, Sigma, P5585), 0.5 mM Oleic acid (OA), 100 μg/mL Cycloheximide (CHX) (MedChemExpress, HY-12320), 100 μM Chloroquine (CQ) (MedChemExpress, HY-17589), 10 μM MG132 (MedChemExpress, HY-12359), 5 μg/mL Actinomycin D (ActD) (MedChemExpress, HY-17559), 1 mM Tauroursodeoxycholic acid (TUDCA) (Selleck, S3654), 1.2 μM Tunicamycin (TM) (Selleck, S7894), 100 nM Thapsigargin (Tha) (Selleck, S7895), 2.5 μM CB-5083 (MedChemExpress, HY-12861) and 100 μM 2-bromopalmitate (2-BP) (Sigma, 21604).

Techniques: Western Blot, Knock-Out, Fluorescence, Staining, Two Tailed Test, Derivative Assay

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: AOPP accumulation correlated with PC and ER stress markers expression in active CD (A–D) Images representative of immunohistochemical staining showed the expression of Lysozyme, AOPP, GRP78 and CHOP, respectively, in intestinal biopsies from 7 patients with CD and 5 healthy controls. Scale bar, 20 μm. Data for the accompanying graphs were generated from immunoreactive scores (IRS). Overall, these results revealed a reduction in Lysozyme (A) with an induction of AOPP (B), GRP78 (C) and CHOP (D). Data were represented by median with range. Student’s t test, ∗∗p < 0.01 versus controls. (E) Pearson correlation and linear analysis exhibited that AOPP accumulation was negatively correlated with Lysozyme expression in intestinal crypts of patients with CD (R = −0.869, p = 0.011). (F and G) Pearson correlation and linear analysis uncovered that increased expression of GRP78 and CHOP were positively correlated with AOPP presence in intestinal crypts of patients with CD (R = 0.793, p = 0.033; R = 0.828, p = 0.022). AOPP, advanced oxidation protein products; CD, Crohn disease; ER, endoplasmic reticulum; GRP78, glucose-regulated protein 78; CHOP, CAAT/enhancer-binding protein (C/EBP) homologous protein.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Expressing, Immunohistochemical staining, Staining, Generated, Binding Assay

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: Chronic administration of AOPP induced PC defects in intestinal crypts of mice (A and B) Representative images of HE staining for PC number quantification and immunohistochemical staining for Lysozyme in intestine sections of mice with or without AOPP treatment. These images were representative of results from each experimental group, each with five mice. Images showed that AOPP challenge reduced the PC number and lysozyme expression in intestinal crypts compared with vehicle-treated mice. Scale bar, 20 μm. (C) Representative Western blotting and densitometric quantification showed the protein level of Lysozyme in isolated intestinal crypts of mice with or without AOPP administration. (D and E) Bar graphs representing qPCR analysis showed the mRNA levels of Lysozyme and cryptdins in isolated intestinal crypts of mice with or without AOPP administration. Relative protein and mRNA levels were normalized by GAPDH, respectively. Data were represented by median with range, n = 5 mice; error bars indicating range. Student’s t test, ∗∗p < 0.01 versus vehicles. Vehicle, PBS treatment for 28 days; AOPP, AOPP treatment for 28 days; qPCR, quantitative real-time polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Staining, Immunohistochemical staining, Expressing, Western Blot, Isolation, Real-time Polymerase Chain Reaction

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: Chronic challenge of AOPP induced ER stress in intestinal crypts of mice (A and B) Representative images of immunohistochemical staining for GRP78 and CHOP in mice with or without AOPP challenge. These images were representative of results from each experimental group, each with five mice. Images showed that AOPP treatment elevated GRP78 and CHOP levels in intestinal crypts compared with vehicle-treated mice. Scale bar, 20 μm. (C) Representative Western blotting and densitometric quantification showed the protein levels of GRP78, CHOP and IRE1α in isolated intestinal crypts of mice with or without AOPP challenge. (D and E) Bar graphs representing qPCR analysis showed the levels of GRP78 and CHOP mRNA in isolated intestinal crypts of mice with or without AOPP challenge. Relative protein and mRNA levels were normalized by GAPDH, respectively. Data were represented by median with range, n = 5 mice; error bars indicating range. Student’s t test, ∗p < 0.05 and ∗∗p < 0.01 versus vehicles.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Immunohistochemical staining, Staining, Western Blot, Isolation

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: AOPP-mediated PC defects depended on ER stress in intestinal crypts of mice (A and B) Representative images of HE staining for PC number quantification and immunohistochemical staining for Lysozyme in intestine sections of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. These images were representative of results from each experimental group, each with five mice. Images reveled that treatment with TUDCA alleviated AOPP-mediated decreasing PC number and lysozyme expression in intestinal crypts compared with AOPP-treated mice. Scale bar, 20 μm. (C) Representative Western blotting and densitometric quantification revealed the protein level of Lysozyme in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. (D and E) Bar graphs representing qPCR analysis revealed the levels of mRNA of Lysozyme and cryptdins in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. Relative protein and mRNA levels were normalized by GAPDH, respectively. Data were represented by median with range, n = 5 mice; error bars indicating range. ANOVA, ∗∗p < 0.01 versus vehicles; # p < 0.05 and ## p < 0.01 versus AOPP-treated mice.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Staining, Immunohistochemical staining, Expressing, Western Blot, Isolation

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: TUDCA inhibited AOPP-resulted ER stress in intestinal crypts of mice (A and B) Representative images of immunohistochemical staining for GRP78 and CHOP in mice that underwent intraperitoneal administration of AOPP with or without TUDCA. These images were representative of results from each experimental group, each with five mice. Images revealed that treatment with TUDCA attenuated AOPP-induced increased GRP78 and CHOP levels in intestinal crypts compared with AOPP-treated mice. Scale bar, 20 μm. (C) Representative Western blotting and densitometric quantification revealed the protein levels of GRP78, CHOP and IRE1α in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. (D and E) Bar graphs representing qPCR analysis revealed the levels of GRP78 and CHOP mRNA in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. Relative protein and mRNA levels were normalized by GAPDH, respectively. Data were represented by median with range, n = 5 mice; error bars indicating range. ANOVA, ∗∗p < 0.01 versus vehicle; # p < 0.05 and ## p < 0.01 versus AOPP-treated mice. AOPP+TUDCA, AOPP treatment plus daily intraperitoneal injection of TUDCA; TUDCA, tauroursodeoxycholic acid, an inhibitor of ER stress.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Immunohistochemical staining, Staining, Western Blot, Isolation, Injection

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: MAM enrichment in AOPP-treated intestinal crypts of mice was mediated by ER stress (A) Representative images of TEM for MAM analysis showed the association of ER with mitochondria in intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. Quantitation of ER length adjacent to mitochondria normalized by mitochondrial perimeter. Scale bar, 2 μm. (B) Representative Western blotting and densitometric quantification showing that the expression of GRP75, VDAC1, Mfn1, and Mfn2 were enhanced in MAM fractions from isolated intestinal crypts of AOPP-treated mice compared with vehicle-treated mice, and the AOPP+TUDCA group demonstrated that administration with TUDCA restored AOPP-resulted enhancive MAM-related protein. PDI served as a loading control. Relative protein levels were expressed as fold induction over vehicle. Data were represented by median with range, n = 5 mice; error bars indicating range. ANOVA, ∗∗p < 0.01 versus vehicles; # p < 0.05 and ## p < 0.01 versus AOPP-treated mice. (C) Representative images of organelle-targeted fluorescence labeling for ER (calnexin) and mitochondria (TOMM20) showed the co-localization of ER and mitochondria, indicating MAM formation in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. Scale bar, 10 μm. MAM, mitochondria-associated ER membranes; Grp75, glucose regulated protein 75; VDAC1, voltage-dependent anion channel 1; Mfn1/2, mitofusin-1/2; TEM, transmission electron microscopy; ER, endoplasmic reticulum; Asterisks, mitochondria; N, nucleus; AG, acidophil granule.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Quantitation Assay, Western Blot, Expressing, Isolation, Control, Fluorescence, Labeling, Transmission Assay, Electron Microscopy

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet: AOPP induced mitochondrial dysfunction in intestinal crypts of mice through ER stress (A) Quantitative luminescence analysis of ATP levels showed the production of ATP in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. (B) Quantitative fluorescence analysis of mitochondrial Ca 2+ levels in isolated intestinal crypts incubating by Rhod-2, a mitochondrial Ca 2+ indicator, showed that AOPP-treated mice increased levels of mitochondrial Ca 2+ concentration compared with vehicle-treated mice, and the TUDCA+AOPP group determined that treatment with TUDCA prevented AOPP-induced mitochondrial Ca 2+ overload. (C) Flow cytometry analysis of MPTP activation showed the opening of MPTP in isolated intestinal crypts of mice that underwent intraperitoneal administration of AOPP with or without TUDCA. Data were represented by median with range, n = 5 mice; error bars indicating range. ANOVA, ∗∗p < 0.01 versus vehicles; # p < 0.05 and ## p < 0.01 versus AOPP-treated mice. ATP, adenosine triphosphate; MPTP, mitochondrial permeability transition pore.

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Quantitative Luminescence, Isolation, Fluorescence, Concentration Assay, Flow Cytometry, Activation Assay, Permeability

Journal: iScience

Article Title: Advanced oxidation protein products induce Paneth cells defects by endoplasmic reticulum stress in Crohn's disease

doi: 10.1016/j.isci.2023.107312

Figure Lengend Snippet:

Article Snippet: AOPPs were prepared in vitro by mixing Rabbit Serum Albumin (Sigma, St Louis, MO) with sodium hypochlorite (Macklin, Shanghai, China) for 30 min, as described previously., , , All mice were randomly divided into three groups (n = 5 in each group) and given intraperitoneal injections of the following: (i) vehicle group: daily injection of phosphate-buffered saline (PBS) for 28 days (pH 7.4); (ii) AOPP group: daily injection of AOPP (50 mg/kg for 28 days); (iii) AOPP plus TUDCA (Selleck, Shanghai, China)-treated group: daily injection of TUDCA (250 mg/kg) for 2 h, before injection of AOPP (50 mg/kg) for 28 days.

Techniques: Recombinant, Lysis, Protein Quantitation, Isolation, ATP Assay, Software

Journal: Nutrients

Article Title: FXR, a Key Regulator of Lipid Metabolism, Is Inhibited by ER Stress-Mediated Activation of JNK and p38 MAPK in Large Yellow Croakers ( Larimichthys crocea ) Fed High Fat Diets

doi: 10.3390/nu13124343

Figure Lengend Snippet: Effects of HFD and ER stress on the expression of FXR in large yellow croaker: ( A , B ) Immunoblots or qRT-PCR assays for FXR and ER stress markers in the liver of large yellow croakers fed HFD diet; ( C – F ) immunoblots or qRT-PCR assays for FXR and ER stress markers in LYCL cells after PA, TG, or TM treatment; ( G ) immunoblots for FXR in LYCL cells after TUDCA treatment. Data are shown as means ± SEMs ( n = 3) and were analyzed using Student’s t -test, * p < 0.05.

Article Snippet: The LYCL cells were treated for 24 h with palmitic acid (PA, 100 μM, Sigma, St. Louis, MO, USA), thapsigargin (TG, 1 μM, MCE, Bloomfield, NJ, USA), tunicamycin (TM, 4 μM, MCE, Bloomfield, NJ, USA) and tauroursodeoxycholate (TUDCA, 0, 50, 100, 150 and 200 μM, MCE, Bloomfield, NJ, USA) to study the effect of ER stress on fxr expression.

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: Heliyon

Article Title: Role of plasmacytoid dendritic cells in vascular dysfunction in mice with renovascular hypertension

doi: 10.1016/j.heliyon.2024.e31799

Figure Lengend Snippet: Mesenteric resistance arteries (MRA) Reactivity Assessment. This figure delineates the evaluation of mesenteric resistance arteries (MRA) reactivity, specifically illustrating the contractility response to sympathetic stimulation (Phenylephrine, PE), as well as endothelium-dependent and -independent relaxation responses to acetylcholine (ACh) and sodium nitroprusside (SNP) across diverse experimental groups. Panel (A, B) represents the control group (C57BL/6J), while panel (C, D) shows C57BL/6J mice subjected to 2K1C surgery. Panel (E, F) displays C57BL/6J mice subjected to 2K1C for four weeks and treated with anti-PDCA-1 for one week. MRA reactivity was thoroughly evaluated in all groups of mice (n = 5) under different conditions, including those with and without ER stress inhibitor (Tauroursodeoxycholic acid: Tudca), autophagy inhibitor (Chloroquine: Chl), and mTOR signaling inhibitor (Rapamycin: Rap). The data presented in this figure provide valuable insights into the vascular response in the context of the experimental manipulations and treatments, shedding light on the potential involvement of ER stress, autophagy, and mTOR signaling pathways in mediating the observed reactivity changes. ns: p > 0.05, *p < 0.05 for 2K1C vs. control and 2K1C + anti-mPDCA-1. One-way ANOVA followed by Tukey's post hoc test was applied. ns: p > 0.05, *p < 0.05 for C57BL/6J vs 2K1C vs 2K1C + anti-PDCA-1.

Article Snippet: To determine the impact of autophagy, ER stress, and mTOR pathways in endothelial cell function, we isolated arteries from each group and incubated them with the following inhibitors: tauroursodeoxycholic acid (Tudca: ER Stress inhibitor) Dose: 10 mM, 30 min, Chem-Impex Int'l INC. Cat#29195, Lot# 002129–20131118; chloroquine (autophagy inhibitor, dose: 10 mM, 30 min, Alfa Aesar, Cat: J64459, Lot: Z23G009); or rapamycin (mTOR inhibitor, dose: 10 mM, 30 min, Abcam, Cat:ab120224, Lot: APN13087-1-1).

Techniques: Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Food Additive Maltodextrin Promotes Endoplasmic Reticulum Stress–Driven Mucus Depletion and Exacerbates Intestinal Inflammation

doi: 10.1016/j.jcmgh.2018.09.002

Figure Lengend Snippet: ER stress inhibition reduces MDX-mediated Ern-2 up-regulation and improves colitis in MDX-fed mice. ( A ) HT29-MTX cells were pretreated with TUDCA (10 μmol/L) or dimethyl sulfoxide (vehicle) and then stimulated with MDX for 1 hour. Ern-2 RNA transcripts were analyzed by real-time PCR. Data are means ± SD of 4 independent experiments. Differences between groups were compared using the 2-tailed Student t test (* P ≤ .05). ( B and C ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Mice were killed on day 45, colonic tissues were isolated, and ( B ) Ern-2 , Ern-1 , and Xbp1s RNA transcripts and ( C ) Muc-2 protein expression were evaluated by real-time PCR and immunofluorescence, respectively. ( B ) Data were generated using 7–10 mice per group from 3 independent experiments. Each point in the graph indicates the RNA expression of the specific transcript in the colon of a single mouse; horizontal bars indicate median value. Differences between groups were compared using the Mann–Whitney U test (* P ≤ .05). ( C ) Pictures are representative of 4 separate experiments in which similar results were obtained. Scale bars : 25 μm. ( D ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Mice were exposed to 1.75% DSS to induce colitis starting from day 35 until death (day 45), and body weight was recorded every other day. Data were generated using 8–9 mice per group from 3 independent experiments and expressed as means ± SEM. Differences among groups were compared using 1-way analysis of variance followed by the Bonferroni post hoc test (** P ≤ .01). ( E and F ) Representative H&E staining of colon sections of mice treated as indicated in panel D and killed on day 45. ( F ) Scatter plot shows the histologic score. Data were generated using 8–9 mice per group from 3 independent experiments and expressed as means ± SD. Differences among groups were compared using 1-way analysis of variance followed by the Bonferroni post hoc test (* P ≤ .05; ** P ≤ .01). ( G and H ) Representative scatter plots showing ( G ) IL1β and ( H ) Lcn-2 RNA expression in colon tissues taken from mice treated as indicated in panel D and killed on day 45. Each point in the graph indicates the RNA expression of the specific transcript in the colon of a single mouse; horizontal bars indicate median value or means ± SD. Data were generated using 8–9 mice per group from 3 independent experiments. Differences among groups were compared using the Kruskal–Wallis test or 1-way analysis of variance followed by the Bonferroni post hoc test (* P ≤ .05). DAPI, 4′,6-diamidino-2-phenylindole; mRNA, messenger RNA.

Article Snippet: In parallel, mice receiving a MDX-enriched diet, together with control mice, were given 250 mg/kg TUDCA (Carbosynth Ltd, Berkshire, UK) intraperitoneally every other day starting from day 21 of diet.

Techniques: Inhibition, Real-time Polymerase Chain Reaction, Injection, Isolation, Expressing, Immunofluorescence, Generated, RNA Expression, MANN-WHITNEY, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Food Additive Maltodextrin Promotes Endoplasmic Reticulum Stress–Driven Mucus Depletion and Exacerbates Intestinal Inflammation

doi: 10.1016/j.jcmgh.2018.09.002

Figure Lengend Snippet: Effect of MDX on mucosa-associated microbiota. ( A and B ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Control mice received drinking water for 45 days. All mice were killed on day 45. ( A ) Relative abundance of phyla and classes are represented for colonic mucosa-associated microbiota. Horizontal bars indicate median value. Data were generated using 4–7 mice per group from 2 independent experiments. Differences among groups were compared using the Kruskal–Wallis test (* P ≤ .05). ( B ) Principal coordinates analysis (PCoA) of the unweighted and weighted UniFrac distance matrix of mucosa-associated bacteria from mice treated as indicated in panel A .

Article Snippet: In parallel, mice receiving a MDX-enriched diet, together with control mice, were given 250 mg/kg TUDCA (Carbosynth Ltd, Berkshire, UK) intraperitoneally every other day starting from day 21 of diet.

Techniques: Injection, Control, Generated, Bacteria