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Incyte corporation ido1 inhibitor 1-mt

<t>IDO1</t> and TDO were expressed in atherosclerotic lesions and codistributed with CD3-positive lymphocytes and CD68-positive macrophages in patients with advanced atherosclerosis. a Pathological changes of the coronary artery were revealed by HE staining (×400). b , c IDO1, TDO and the lymphocyte marker CD3 were detected in atherosclerotic lesions by immunostaining analysis (×400). d , e IDO1, TDO and the monocyte marker CD68 were detected in atherosclerotic lesions by immunostaining analysis (×400). The top panel in b – e exhibits an enlarged view of the representative region in the bottom panel

Ido1 Inhibitor 1 Mt, supplied by Incyte corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ido1 inhibitor 1-mt - by Bioz Stars, 2026-08

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1) Product Images from "The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis"

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

Journal: Signal Transduction and Targeted Therapy

doi: 10.1038/s41392-019-0058-5

IDO1 and TDO were expressed in atherosclerotic lesions and codistributed with CD3-positive lymphocytes and CD68-positive macrophages in patients with advanced atherosclerosis. a Pathological changes of the coronary artery were revealed by HE staining (×400). b , c IDO1, TDO and the lymphocyte marker CD3 were detected in atherosclerotic lesions by immunostaining analysis (×400). d , e IDO1, TDO and the monocyte marker CD68 were detected in atherosclerotic lesions by immunostaining analysis (×400). The top panel in b – e exhibits an enlarged view of the representative region in the bottom panel

Figure Legend Snippet: IDO1 and TDO were expressed in atherosclerotic lesions and codistributed with CD3-positive lymphocytes and CD68-positive macrophages in patients with advanced atherosclerosis. a Pathological changes of the coronary artery were revealed by HE staining (×400). b , c IDO1, TDO and the lymphocyte marker CD3 were detected in atherosclerotic lesions by immunostaining analysis (×400). d , e IDO1, TDO and the monocyte marker CD68 were detected in atherosclerotic lesions by immunostaining analysis (×400). The top panel in b – e exhibits an enlarged view of the representative region in the bottom panel

Techniques Used: Staining, Marker, Immunostaining

The expression and activity of IDO1 and TDO in atherosclerotic blood samples. Serum concentrations of tryptophan ( a ) and kynurenine ( b ) and the kynurenine to tryptophan ratio Kyn/Trp ( c ) in blood samples of atherosclerotic patients with different histological classification grades ((−): n = 10, I: n = 16, II: n = 18, III: n = 7). d , e IDO1 and TDO protein levels were determined by western blot assays. f Associations between the expression of IDO1 and TDO and the concentration of Trp ( r = −0.44028, * p < 0.05; r = −0.45339, * p < 0.05). g IDO1 and TDO mRNA expression was determined by real-time PCR ((−): n = 6, I/II: n = 12, III: n = 4). Data were based on at least three independent experiments. * p < 0.05

Figure Legend Snippet: The expression and activity of IDO1 and TDO in atherosclerotic blood samples. Serum concentrations of tryptophan ( a ) and kynurenine ( b ) and the kynurenine to tryptophan ratio Kyn/Trp ( c ) in blood samples of atherosclerotic patients with different histological classification grades ((−): n = 10, I: n = 16, II: n = 18, III: n = 7). d , e IDO1 and TDO protein levels were determined by western blot assays. f Associations between the expression of IDO1 and TDO and the concentration of Trp ( r = −0.44028, * p < 0.05; r = −0.45339, * p < 0.05). g IDO1 and TDO mRNA expression was determined by real-time PCR ((−): n = 6, I/II: n = 12, III: n = 4). Data were based on at least three independent experiments. * p < 0.05

Techniques Used: Expressing, Activity Assay, Western Blot, Concentration Assay, Real-time Polymerase Chain Reaction

oxLDL induced IDO1 expression via the PI3K/Akt/NF-κB pathway in THP-M. a oxLDL induced the production of inflammatory factors. The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. b THP-M were challenged with 25 mg/L oxLDL for 12, 24, and 48 h, and IDO1 expression was detected by western blot analysis. c THP-M were treated with oxLDL for 24 h at 0, 25, 50 and 100 mg/L, and IDO1 expression was detected by western blot analysis. d The expression of PI3K/Akt/NF-κB pathway-related proteins in THP-M was detected by western blot analysis. THP-M were treated with oxLDL (25 mg/L) for 24 h. e The expression levels of pAkt, pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the Akt inhibitor LY294002 (5 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h. Data were based on at least three independent experiments. * p < 0.05. f The expression levels of pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the NF-κB inhibitor CAPE (50 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h

Figure Legend Snippet: oxLDL induced IDO1 expression via the PI3K/Akt/NF-κB pathway in THP-M. a oxLDL induced the production of inflammatory factors. The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. b THP-M were challenged with 25 mg/L oxLDL for 12, 24, and 48 h, and IDO1 expression was detected by western blot analysis. c THP-M were treated with oxLDL for 24 h at 0, 25, 50 and 100 mg/L, and IDO1 expression was detected by western blot analysis. d The expression of PI3K/Akt/NF-κB pathway-related proteins in THP-M was detected by western blot analysis. THP-M were treated with oxLDL (25 mg/L) for 24 h. e The expression levels of pAkt, pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the Akt inhibitor LY294002 (5 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h. Data were based on at least three independent experiments. * p < 0.05. f The expression levels of pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the NF-κB inhibitor CAPE (50 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Before and after oxLDL treatment, the promoting effects of IFN-γ on IDO1 expression and the degree of foaming of THP-M. a Induction of IDO1 by different treatments was detected by immunofluorescence (×200). b The degree of foaming was detected by Oil red O staining (×400). c Quantitative analysis of Oil red O intensity (Oil red O staining area/cell area and Oil red O staining particles per cell). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+ oxLDL and oxLDL+IFN-γ, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Figure Legend Snippet: Before and after oxLDL treatment, the promoting effects of IFN-γ on IDO1 expression and the degree of foaming of THP-M. a Induction of IDO1 by different treatments was detected by immunofluorescence (×200). b The degree of foaming was detected by Oil red O staining (×400). c Quantitative analysis of Oil red O intensity (Oil red O staining area/cell area and Oil red O staining particles per cell). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+ oxLDL and oxLDL+IFN-γ, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Techniques Used: Expressing, Foaming, Immunofluorescence, Staining

Before and after oxLDL treatment, the IFN-γ-mediated promotion of THP-M foaming was reversed by IDO1 inhibition. a , b IDO1 activity (Kyn/Trp) was detected by HPLC. c , d The results of Oil red O staining (×400). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05

Figure Legend Snippet: Before and after oxLDL treatment, the IFN-γ-mediated promotion of THP-M foaming was reversed by IDO1 inhibition. a , b IDO1 activity (Kyn/Trp) was detected by HPLC. c , d The results of Oil red O staining (×400). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05

Techniques Used: Foaming, Inhibition, Activity Assay, Staining

Before and after oxLDL treatment, IFN-γ-induced IDO1 exhibited different degrees of promotion on cell apoptosis and inflammatory factor production. a The degree of cell apoptosis was evaluated by flow cytometry. PI for nuclear staining and Annexin V-FITC for cytomembrane staining. The UL (upper left) quadrant represents cellular debris and damaged cells. The UR (upper right) and LR (lower right) quadrants represent apoptotic cells in the late or early stages. The LL (lower left) quadrant shows the normal cells. b Quantitative analysis of cell apoptosis. The total cell apoptosis is the sum of the percentage of late cell and early cell apoptosis. c Western blot analysis of caspase-3 expression in THP-M. d The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Figure Legend Snippet: Before and after oxLDL treatment, IFN-γ-induced IDO1 exhibited different degrees of promotion on cell apoptosis and inflammatory factor production. a The degree of cell apoptosis was evaluated by flow cytometry. PI for nuclear staining and Annexin V-FITC for cytomembrane staining. The UL (upper left) quadrant represents cellular debris and damaged cells. The UR (upper right) and LR (lower right) quadrants represent apoptotic cells in the late or early stages. The LL (lower left) quadrant shows the normal cells. b Quantitative analysis of cell apoptosis. The total cell apoptosis is the sum of the percentage of late cell and early cell apoptosis. c Western blot analysis of caspase-3 expression in THP-M. d The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Techniques Used: Flow Cytometry, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Before and after oxLDL treatment, the IFN-γ-mediated promotion of cell apoptosis and inflammatory factor production were reversed by IDO1 inhibition. a Western blot analysis of caspase-3 expression in THP-M. b The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. The designation of different treatments, such as IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Figure Legend Snippet: Before and after oxLDL treatment, the IFN-γ-mediated promotion of cell apoptosis and inflammatory factor production were reversed by IDO1 inhibition. a Western blot analysis of caspase-3 expression in THP-M. b The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. The designation of different treatments, such as IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Techniques Used: Inhibition, Western Blot, Expressing, Real-time Polymerase Chain Reaction

The IDO1 inhibitor 1-MT could ameliorate the development of atherogenesis in high-fat diet (HFD)-fed ApoE −/− mice. Eight-week-old WT and ApoE −/− mice were used and killed at 12 weeks of age. a Representative photomicrographs of aortic roots from 12-week-old mice of different groups stained with Oil red O. b The atherosclerotic plaque area/total aortic sinus area ratio. Atherosclerotic lesion areas were measured as the mean sizes of multiple plaques located in 5 sections in each mouse. c IDO1 activity analyzed by HPLC (WT group: n = 6, CMC group: n = 4, 1-MT group: n = 4). d Total cholesterol (TC). e High-density lipoprotein cholesterol (HDL-C). f Low-density lipoprotein cholesterol (LDL-C). n = 4. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Figure Legend Snippet: The IDO1 inhibitor 1-MT could ameliorate the development of atherogenesis in high-fat diet (HFD)-fed ApoE −/− mice. Eight-week-old WT and ApoE −/− mice were used and killed at 12 weeks of age. a Representative photomicrographs of aortic roots from 12-week-old mice of different groups stained with Oil red O. b The atherosclerotic plaque area/total aortic sinus area ratio. Atherosclerotic lesion areas were measured as the mean sizes of multiple plaques located in 5 sections in each mouse. c IDO1 activity analyzed by HPLC (WT group: n = 6, CMC group: n = 4, 1-MT group: n = 4). d Total cholesterol (TC). e High-density lipoprotein cholesterol (HDL-C). f Low-density lipoprotein cholesterol (LDL-C). n = 4. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Techniques Used: Staining, Activity Assay

Figure Legend Snippet: Primers used for real-time PCR

Techniques Used:

Image Search Results

Fig. 1. APAP-induced acute liver failure involved lipid peroxidation, excess nitrative stress and iron up-regulation, accompanied by increased expression of IDO1. (A) Representative images of surface morphology and color of mice livers in different groups. (B) Hematoxylin-eosin (H&E) staining and TUNEL staining of liver paraffin sections were shown from control group and 300 mg/kg APAP treated group (dotted lines in white showed damaged areas). (C) Serum concentrations of AST/ALT between control group and APAP group. (D) The RNS production after acute liver failure stained positive for NP3 probe (blue). PI nucleic counterstain appeared in red. (E)TGF-βr and IFN-γ protein expression. GAPDH served as a loading control respectively. (F-H) The hepatic levels of SOD activity, GSH and MDA in mice of control group and APAP group were determined.(I)Iron concentration (including total iron, ferrous iron and ferric iron) in control group and APAP-treated mice was detected. (J) Representative electron microscope images of morphological structure of mitochondria in hepatocytes (arrows in white showed lesions). (K) Western blot analysis reveals increased TfR and Tf after 24 h of APAP injection in mice. GAPDH served as a loading control respectively. (L) Western blot and immunohistochemical representative images of IDO1 in control group and APAP group. GAPDH served as a loading control respectively. The density of protein, the fluorescence intensity and the area of immunohistochemistry staining was measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 1. APAP-induced acute liver failure involved lipid peroxidation, excess nitrative stress and iron up-regulation, accompanied by increased expression of IDO1. (A) Representative images of surface morphology and color of mice livers in different groups. (B) Hematoxylin-eosin (H&E) staining and TUNEL staining of liver paraffin sections were shown from control group and 300 mg/kg APAP treated group (dotted lines in white showed damaged areas). (C) Serum concentrations of AST/ALT between control group and APAP group. (D) The RNS production after acute liver failure stained positive for NP3 probe (blue). PI nucleic counterstain appeared in red. (E)TGF-βr and IFN-γ protein expression. GAPDH served as a loading control respectively. (F-H) The hepatic levels of SOD activity, GSH and MDA in mice of control group and APAP group were determined.(I)Iron concentration (including total iron, ferrous iron and ferric iron) in control group and APAP-treated mice was detected. (J) Representative electron microscope images of morphological structure of mitochondria in hepatocytes (arrows in white showed lesions). (K) Western blot analysis reveals increased TfR and Tf after 24 h of APAP injection in mice. GAPDH served as a loading control respectively. (L) Western blot and immunohistochemical representative images of IDO1 in control group and APAP group. GAPDH served as a loading control respectively. The density of protein, the fluorescence intensity and the area of immunohistochemistry staining was measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Expressing, Staining, TUNEL Assay, Control, Activity Assay, Concentration Assay, Microscopy, Western Blot, Injection, Immunohistochemical staining, Fluorescence, Immunohistochemistry, Software

Fig. 3. IDO1 deficiency enhanced antioxidant capacity and suppressed lipid peroxidation in ALF. (A) Serum concentrations of AST and ALT were measured. (B, C) Hematoxylin–eosin (H&E) stained and TUNEL stained sections of the livers were shown (dotted lines in white showed damaged areas). (D) Western blot analysis relative density ratios of iNOS, COX-2 and Cyto-C expression in different groups of WT and IDO1−/−mice livers at 24 h after APAP administration. GAPDH served as a loading control. (E) Immunohistochemical representative images of 4-HNE in different groups. (F, G) MDA and GSH levels in mice of different indicated groups were determined. (H) Representative ROS probe detection pictures in LO2 cells were shown. Quantitative analysis in above panel. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 3. IDO1 deficiency enhanced antioxidant capacity and suppressed lipid peroxidation in ALF. (A) Serum concentrations of AST and ALT were measured. (B, C) Hematoxylin–eosin (H&E) stained and TUNEL stained sections of the livers were shown (dotted lines in white showed damaged areas). (D) Western blot analysis relative density ratios of iNOS, COX-2 and Cyto-C expression in different groups of WT and IDO1−/−mice livers at 24 h after APAP administration. GAPDH served as a loading control. (E) Immunohistochemical representative images of 4-HNE in different groups. (F, G) MDA and GSH levels in mice of different indicated groups were determined. (H) Representative ROS probe detection pictures in LO2 cells were shown. Quantitative analysis in above panel. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Staining, TUNEL Assay, Western Blot, Expressing, Control, Immunohistochemical staining, Fluorescence, Software

Fig. 4. IDO1 deficiency performed stronger resistance to excess nitrative stress. (A) Immunofluorescence staining of 3-NT (red) in livers in vehicle, APAP- treated, IDO1−/−vehicle and IDO1−/−APAP treated mice. (B) Fluorescence micrographs of ONOO- probe detection (blue) of livers. Nuclei were stained with PI (red). The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 4. IDO1 deficiency performed stronger resistance to excess nitrative stress. (A) Immunofluorescence staining of 3-NT (red) in livers in vehicle, APAP- treated, IDO1−/−vehicle and IDO1−/−APAP treated mice. (B) Fluorescence micrographs of ONOO- probe detection (blue) of livers. Nuclei were stained with PI (red). The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Immunofluorescence, Staining, Fluorescence, Software

Fig. 5. Inhibition of IDO1 contributed to hepatocytes production and inflammatory cells reduction. (A) Immunofluorescence staining of F4/80 in livers. (B, C) The serum levels of IL-6 and TGF-β in mice of different indicated groups were detected. (D) Schematic illustration of the zebrafish drug toxicity experiment. Zebrafish were cultured for 48 h after exposed to egg water or medias for 48 h before imaging respectively. (E) Survival proportions of zebrafish from 0 to 48 h post exposure to egg water, or APAP (5 mM, 10 mM). Zebrafish survival status were observed at 0, 12, 24, 36 and 48 hpe, n=30 per group. (F) Quantitative analysis of hepatocytes fluorescence in zebrafish. Figures are magnified as 64 × . n=8-10 per group. (G, H) Quantitative analysis of systemic neutrophils and macrophages fluorescence in zebrafish. n=8-10 per group. Zebrafish morphology imaged by microscope. Figures are magnified as 32 × (whole fish) and 64 × (tail). The fluo rescence intensity was measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 5. Inhibition of IDO1 contributed to hepatocytes production and inflammatory cells reduction. (A) Immunofluorescence staining of F4/80 in livers. (B, C) The serum levels of IL-6 and TGF-β in mice of different indicated groups were detected. (D) Schematic illustration of the zebrafish drug toxicity experiment. Zebrafish were cultured for 48 h after exposed to egg water or medias for 48 h before imaging respectively. (E) Survival proportions of zebrafish from 0 to 48 h post exposure to egg water, or APAP (5 mM, 10 mM). Zebrafish survival status were observed at 0, 12, 24, 36 and 48 hpe, n=30 per group. (F) Quantitative analysis of hepatocytes fluorescence in zebrafish. Figures are magnified as 64 × . n=8-10 per group. (G, H) Quantitative analysis of systemic neutrophils and macrophages fluorescence in zebrafish. n=8-10 per group. Zebrafish morphology imaged by microscope. Figures are magnified as 32 × (whole fish) and 64 × (tail). The fluo rescence intensity was measured using image J software. Statistical analysis demonstrated the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Inhibition, Immunofluorescence, Staining, Cell Culture, Imaging, Fluorescence, Microscopy, Software

Fig. 6. IDO1 deficiency reduced iron accumulation via suppressing Tf-TfR axis. (A) Iron accumulation in different groups. (B) Immunoblotting results showed that comparing with APAP-treated WT mice, APAP-treated IDO1−/−mice inhibited Tf and TfR expression obviously. GAPDH served as a loading control. Quantitative analysis in right panel. (C, D) Immunofluorescence staining of Tf and TfR in liver tissue of both WT mice and IDO1−/−mice. Quantitative analysis in right panel respectively. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis showed the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 6. IDO1 deficiency reduced iron accumulation via suppressing Tf-TfR axis. (A) Iron accumulation in different groups. (B) Immunoblotting results showed that comparing with APAP-treated WT mice, APAP-treated IDO1−/−mice inhibited Tf and TfR expression obviously. GAPDH served as a loading control. Quantitative analysis in right panel. (C, D) Immunofluorescence staining of Tf and TfR in liver tissue of both WT mice and IDO1−/−mice. Quantitative analysis in right panel respectively. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis showed the mean + SEM, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Staining, Fluorescence, Software

Fig. 7. Exhaustion of macrophages alleviated liver injury by reducing iron accumulation in hepatocytes. (A) Relative number of cells expression of CD11b and CD71 were showed by Flow cytometry. (B) Representative Immunofluorescence staining of FPN1 in RAW264.7 macrophages treated with APAP (5 mM) for 8 h. (C) Western blot showed the expression of TfR, FPN1 and IDO1 protein after 24 h of incubation with Erastin (40 μM or 80 μM) in RAW264.7 cells. (D) Representative Immunofluorescence staining of F4/80 (green) expression in mice liver. Nuclei were stained with DAPI (blue). (E, F) The serum levels of inflammatory IL-6 and MCP- 1 were detected. (G–I) Comparison of liver tissue homogenate of iron concentration, MDA and SOD level of different indicated groups were determined. (J) Immunoblotting results showed that comparing with APAP-treated WT mice, Gdcl3 treated mice inhibited TfR expression obviously. GAPDH served as a loading control. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis showed the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Free radical biology & medicine

Article Title: Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation.

doi: 10.1016/j.freeradbiomed.2021.07.008

Figure Lengend Snippet: Fig. 7. Exhaustion of macrophages alleviated liver injury by reducing iron accumulation in hepatocytes. (A) Relative number of cells expression of CD11b and CD71 were showed by Flow cytometry. (B) Representative Immunofluorescence staining of FPN1 in RAW264.7 macrophages treated with APAP (5 mM) for 8 h. (C) Western blot showed the expression of TfR, FPN1 and IDO1 protein after 24 h of incubation with Erastin (40 μM or 80 μM) in RAW264.7 cells. (D) Representative Immunofluorescence staining of F4/80 (green) expression in mice liver. Nuclei were stained with DAPI (blue). (E, F) The serum levels of inflammatory IL-6 and MCP- 1 were detected. (G–I) Comparison of liver tissue homogenate of iron concentration, MDA and SOD level of different indicated groups were determined. (J) Immunoblotting results showed that comparing with APAP-treated WT mice, Gdcl3 treated mice inhibited TfR expression obviously. GAPDH served as a loading control. The density of protein and the fluorescence intensity were measured using image J software. Statistical analysis showed the mean + SEM, *p<0.05, **p<0.01, ***p<0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The IDO1 inhibitor 1-MT (1-methyl-tryptophan, Selleck) was solubled in dimethyl sulfoxide (DMSO) and diluted to 50 μM with filtered egg water.

Techniques: Expressing, Flow Cytometry, Immunofluorescence, Staining, Western Blot, Incubation, Comparison, Concentration Assay, Control, Fluorescence, Software

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: IDO1 and TDO were expressed in atherosclerotic lesions and codistributed with CD3-positive lymphocytes and CD68-positive macrophages in patients with advanced atherosclerosis. a Pathological changes of the coronary artery were revealed by HE staining (×400). b , c IDO1, TDO and the lymphocyte marker CD3 were detected in atherosclerotic lesions by immunostaining analysis (×400). d , e IDO1, TDO and the monocyte marker CD68 were detected in atherosclerotic lesions by immunostaining analysis (×400). The top panel in b – e exhibits an enlarged view of the representative region in the bottom panel

Article Snippet: We found that IFN-γ-induced IDO1 could promote the transformation of THP-M to foam cells, and two classical IDO1 inhibitors, Incyte and 1-MT, could inhibit IFN-γ-induced IDO1 (ref. ) and weaken the foaming.

Techniques: Staining, Marker, Immunostaining

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: The expression and activity of IDO1 and TDO in atherosclerotic blood samples. Serum concentrations of tryptophan ( a ) and kynurenine ( b ) and the kynurenine to tryptophan ratio Kyn/Trp ( c ) in blood samples of atherosclerotic patients with different histological classification grades ((−): n = 10, I: n = 16, II: n = 18, III: n = 7). d , e IDO1 and TDO protein levels were determined by western blot assays. f Associations between the expression of IDO1 and TDO and the concentration of Trp ( r = −0.44028, * p < 0.05; r = −0.45339, * p < 0.05). g IDO1 and TDO mRNA expression was determined by real-time PCR ((−): n = 6, I/II: n = 12, III: n = 4). Data were based on at least three independent experiments. * p < 0.05

Techniques: Expressing, Activity Assay, Western Blot, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: oxLDL induced IDO1 expression via the PI3K/Akt/NF-κB pathway in THP-M. a oxLDL induced the production of inflammatory factors. The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. b THP-M were challenged with 25 mg/L oxLDL for 12, 24, and 48 h, and IDO1 expression was detected by western blot analysis. c THP-M were treated with oxLDL for 24 h at 0, 25, 50 and 100 mg/L, and IDO1 expression was detected by western blot analysis. d The expression of PI3K/Akt/NF-κB pathway-related proteins in THP-M was detected by western blot analysis. THP-M were treated with oxLDL (25 mg/L) for 24 h. e The expression levels of pAkt, pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the Akt inhibitor LY294002 (5 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h. Data were based on at least three independent experiments. * p < 0.05. f The expression levels of pp65, IDO1 and β-actin were detected by western blots. THP-M were pretreated with the NF-κB inhibitor CAPE (50 μM) for 24 h and then treated with oxLDL (25 mg/L) for 24 h

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: Before and after oxLDL treatment, the promoting effects of IFN-γ on IDO1 expression and the degree of foaming of THP-M. a Induction of IDO1 by different treatments was detected by immunofluorescence (×200). b The degree of foaming was detected by Oil red O staining (×400). c Quantitative analysis of Oil red O intensity (Oil red O staining area/cell area and Oil red O staining particles per cell). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+ oxLDL and oxLDL+IFN-γ, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Techniques: Expressing, Foaming, Immunofluorescence, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: Before and after oxLDL treatment, the IFN-γ-mediated promotion of THP-M foaming was reversed by IDO1 inhibition. a , b IDO1 activity (Kyn/Trp) was detected by HPLC. c , d The results of Oil red O staining (×400). The designation of different treatments, such as oxLDL, IFN-γ, IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05

Techniques: Foaming, Inhibition, Activity Assay, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: Before and after oxLDL treatment, IFN-γ-induced IDO1 exhibited different degrees of promotion on cell apoptosis and inflammatory factor production. a The degree of cell apoptosis was evaluated by flow cytometry. PI for nuclear staining and Annexin V-FITC for cytomembrane staining. The UL (upper left) quadrant represents cellular debris and damaged cells. The UR (upper right) and LR (lower right) quadrants represent apoptotic cells in the late or early stages. The LL (lower left) quadrant shows the normal cells. b Quantitative analysis of cell apoptosis. The total cell apoptosis is the sum of the percentage of late cell and early cell apoptosis. c Western blot analysis of caspase-3 expression in THP-M. d The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Techniques: Flow Cytometry, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: Before and after oxLDL treatment, the IFN-γ-mediated promotion of cell apoptosis and inflammatory factor production were reversed by IDO1 inhibition. a Western blot analysis of caspase-3 expression in THP-M. b The mRNA levels of IL-10, MCP-1, IL-1β and IL-8 were measured by real-time PCR. The designation of different treatments, such as IFN-γ+oxLDL, oxLDL+IFN-γ, IFN-γ+Incyte+oxLDL, oxLDL+IFN-γ+Incyte, IFN-γ+1-MT+oxLDL and oxLDL+IFN-γ+1-MT, was described in the Materials and Methods. Data were based on at least three independent experiments. * p < 0.05, ** p < 0.01

Techniques: Inhibition, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: The IDO1 inhibitor 1-MT could ameliorate the development of atherogenesis in high-fat diet (HFD)-fed ApoE −/− mice. Eight-week-old WT and ApoE −/− mice were used and killed at 12 weeks of age. a Representative photomicrographs of aortic roots from 12-week-old mice of different groups stained with Oil red O. b The atherosclerotic plaque area/total aortic sinus area ratio. Atherosclerotic lesion areas were measured as the mean sizes of multiple plaques located in 5 sections in each mouse. c IDO1 activity analyzed by HPLC (WT group: n = 6, CMC group: n = 4, 1-MT group: n = 4). d Total cholesterol (TC). e High-density lipoprotein cholesterol (HDL-C). f Low-density lipoprotein cholesterol (LDL-C). n = 4. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Techniques: Staining, Activity Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: The proatherosclerotic function of indoleamine 2, 3-dioxygenase 1 in the developmental stage of atherosclerosis

doi: 10.1038/s41392-019-0058-5

Figure Lengend Snippet: Primers used for real-time PCR

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