acid 1 benzyl ester  (Chem Impex International)

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane‐induced preconditioning improves outcomes after cerebral ischemia and upregulates ATF5 and enhances mitochondria function. A) Experimental timeline. B) Representative 2,3,5‐triphenyltetrazolium chloride (TTC)‐stained coronal brain sections showing infarct areas (white) in sham, control, and sevoflurane‐treated groups with different exposure durations (0.5, 1, 2, 3 h). C) Quantification of cortical infarct volume and neurological scores (n = 4–9 per group). D) Western blot analysis of cortical samples obtained 24 h after sevoflurane exposure (n = 5 per group). E) Top: Mitochondrial OCR measured in cortical mitochondria after sequential administration of ADP, oligomycin, CCCP, and antimycin (arrows indicate time of injection). Bottom: Quantification of OCR after subtracting non‐mitochondrial respiration (n = 6 per group). F) TOMM40 and NeuN immunofluorescence staining in coronal brain sections from control and sevoflurane‐treated mice. High magnification images (right panels) show increased TOMM40 signals (white arrows) in the penumbra after sevoflurane preconditioning. G) Differential expression analysis of 83 mitochondria genes in non‐infarct (top) and penumbra (bottom) cortical areas. Mitochondria function‐related genes are listed in Table (Supporting Information). Volcano plots show the number of significantly changed genes (red) in each region after sevoflurane preconditioning and MCAO (n = 4 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Staining, Control, Western Blot, Injection, Immunofluorescence, Quantitative Proteomics

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane induces mitochondrial dysfunction and fails to induce preconditioning in Atf5 ‐cKO mice. A) Representative IHC images of cultured primary neurons (n = 18 per group). B) Representative IHC images of anterior cingulate cortex layer V‐VI of Atf5 ‐cKO mice showing the colocalization of CAMKII (green) and ATF5 (red) immunostaining and DAPI‐stained nuclei (blue). White arrows indicate decreased, but remnant, ATF5 expression in excitatory neurons. C) Representative TTC‐stained images of MCAO model Sham, WT, and Atf5 ‐cKO mice after sevoflurane exposure. D) Summary data showing decreased cerebral infarct size and improved neurological score in WT mice but not Atf5 ‐cKO mice (n = 5–7 per group). E) mRNA expression levels of UPR mt ‐related genes in the cortices of WT mice after exposure to sevoflurane for 6 h (n = 4–9 per group). F) mRNA expression levels of UPR mt ‐related genes in the cortices of Atf5 ‐cKO mice after exposure to sevoflurane for 6 h (n = 5 per group). G,H) Western blot analysis of cortical samples from Atf5 ‐cKO mice, obtained 24 h after sevoflurane exposure (n = 4–5 per group). I) Left: Mitochondrial function, measured as mitochondrial OCR, determined by assessing respiration of mitochondria isolated from the cortices of mice in control and sevoflurane groups (n = 5 per group). ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. Right: Quantification of OCR after excluding non‐mitochondrial respiration (n = 5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Cell Culture, Immunostaining, Staining, Expressing, Western Blot, Isolation, Control

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane induces mitochondrial dysfunction and fails to induce preconditioning in Gdf15 ‐KO mice. A) Representative TTC‐stained images of MCAO model WT and Gdf15 ‐KO mice after sevoflurane (Sevo) exposure. B) Summary data showing decreased cerebral infarct volume (n = 4–5 per group). C) Volcano plots showing mitochondrial energy metabolism‐related genes whose levels were increased in the cerebral cortex of WT and Gdf15 ‐KO mice 6 h after exposure to sevoflurane (n = 3–5 per group). D) Mitochondrial function, measured as mitochondrial OCR, was determined by assessing the respiration of mitochondria isolated from the cortices of mice in control and sevoflurane groups. ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. E) Quantification of OCR after excluding non‐mitochondrial respiration (n = 3–5 per group). F,G) Western blot analysis of cortical samples obtained from WT and Gdf15 ‐KO mice 24 h after sevoflurane exposure (n = 4–5 per group). H–J) Serum GDF15 protein levels prior to and 3, 6, and 24 h after exposure to sevoflurane in C57BL/6J, WT, and Atf5 ‐cKO mice. K) Comparison of serum GDF15 directly after sevoflurane exposure in C57BL/6J, WT, and Atf5 ‐cKO mice (n = 3–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Staining, Isolation, Control, Western Blot, Comparison

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: ATF5 overexpression in excitatory neurons upregulates GDF15 and enhances mitochondrial function. A) Representative IHC images confirming pAAV‐hSyn‐DIO‐ATF5 expression after stereotaxic injection together with pAAV‐hSyn‐DIO‐EGFP into the cortex of Emx1 cre/+ mice. B) Western blot analysis of cortical samples surrounding the injection site, performed 4 weeks after AAV injection (n = 5 per group). C) Volcano plots showing mitochondrial energy metabolism‐related genes (including Gdf15 ) whose levels were increased around the injection region (n = 5 per group). D) Left: Mitochondrial function, measured as mitochondrial OCR, determined by assessing respiration of mitochondria isolated from the cortex region injected with control or ATF5 virus (n = 5 per group). ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. Right: Quantification of OCR after excluding non‐mitochondrial respiration (n = 5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Over Expression, Expressing, Injection, Western Blot, Isolation, Control, Virus

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: ATF5 overexpression in excitatory neurons provides neuroprotection against ischemic injury. A) Upper panel: Representative image of EGFP (green) and ATF5 (red) expression in the cortex 4 weeks after intravenous injection of pAAV‐hSyn‐DIO‐EGFP. Lower panel: Representative image of CAMKII (green) and ATF5 (red) expression in the cortex 4 weeks after intravenous injection of pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice. B,C) Western blot analysis of ATF5 protein levels in the cortex after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice (n = 5 per group). D) RT‐qPCR analysis of Gdf15 mRNA expression in the cortex after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice (n = 5 per group). E) Serum GDF15 protein levels in Emx1 cre/+ mice 4 weeks after injection with pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 (n = 5 per group). F) Representative TTC‐stained coronal brain sections after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice. G) Quantification of cortical infarct volume and neurological score 24 h after MCAO (n = 4–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Over Expression, Expressing, Injection, Western Blot, Quantitative RT-PCR, Staining

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane‐induced activation of ATF5‐dependent UPR mt , GDF15 expression, and mitochondrial protection are absent in cerebral cortices of aged mice. A) Western blot analysis of ATF5 protein levels in the cortex of mice at 2‐, 8‐, 14‐, and 20‐month of age (n = 5 per group). B) Gdf15 mRNA expression in the cortex at 2‐, 8‐, 14‐, and 20‐ months, determined by RT‐qPCR and normalized to β‐actin (n = 3–4 per group). C) Mitochondrial OCR in isolated cortical mitochondria from 2‐, 8‐, 14‐, and 20‐month‐old mice. Left: Representative OCR traces in response to sequential addition of ADP, oligomycin, CCCP, and antimycin. Right: Quantification of basal, state 3, state 4o, and state 3u respiration. D) Volcano plot showing differential expression of 83 mitochondrial energy metabolism‐related genes in the cortex of 20‐month‐old versus 2‐month‐old mice. Red dots, upregulated genes; blue dots, downregulated genes; black dots, non‐significant changes. E) Western blot analysis of proteins in cortical samples from aged mice obtained 24 h after sevoflurane exposure (n = 5 per group). F) Volcano plot showing changes in mitochondrial energy metabolism‐related genes in the cortex of aged mice 6 h after sevoflurane exposure (n = 3–4 per group). G) Left: Mitochondrial function assessed by OCR in isolated cortical mitochondria from aged mice in control and sevoflurane‐treated groups (n = 4–5 per group). Sequential additions of ADP, oligomycin, CCCP, and antimycin are indicated by arrows. Right: Quantification of OCR, excluding non‐mitochondrial respiration (n = 4–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Activation Assay, Expressing, Western Blot, Quantitative RT-PCR, Isolation, Quantitative Proteomics, Control

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane‐induced preconditioning improves outcomes after cerebral ischemia and upregulates ATF5 and enhances mitochondria function. A) Experimental timeline. B) Representative 2,3,5‐triphenyltetrazolium chloride (TTC)‐stained coronal brain sections showing infarct areas (white) in sham, control, and sevoflurane‐treated groups with different exposure durations (0.5, 1, 2, 3 h). C) Quantification of cortical infarct volume and neurological scores (n = 4–9 per group). D) Western blot analysis of cortical samples obtained 24 h after sevoflurane exposure (n = 5 per group). E) Top: Mitochondrial OCR measured in cortical mitochondria after sequential administration of ADP, oligomycin, CCCP, and antimycin (arrows indicate time of injection). Bottom: Quantification of OCR after subtracting non‐mitochondrial respiration (n = 6 per group). F) TOMM40 and NeuN immunofluorescence staining in coronal brain sections from control and sevoflurane‐treated mice. High magnification images (right panels) show increased TOMM40 signals (white arrows) in the penumbra after sevoflurane preconditioning. G) Differential expression analysis of 83 mitochondria genes in non‐infarct (top) and penumbra (bottom) cortical areas. Mitochondria function‐related genes are listed in Table (Supporting Information). Volcano plots show the number of significantly changed genes (red) in each region after sevoflurane preconditioning and MCAO (n = 4 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Staining, Control, Western Blot, Injection, Immunofluorescence, Quantitative Proteomics

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane induces mitochondrial dysfunction and fails to induce preconditioning in Atf5 ‐cKO mice. A) Representative IHC images of cultured primary neurons (n = 18 per group). B) Representative IHC images of anterior cingulate cortex layer V‐VI of Atf5 ‐cKO mice showing the colocalization of CAMKII (green) and ATF5 (red) immunostaining and DAPI‐stained nuclei (blue). White arrows indicate decreased, but remnant, ATF5 expression in excitatory neurons. C) Representative TTC‐stained images of MCAO model Sham, WT, and Atf5 ‐cKO mice after sevoflurane exposure. D) Summary data showing decreased cerebral infarct size and improved neurological score in WT mice but not Atf5 ‐cKO mice (n = 5–7 per group). E) mRNA expression levels of UPR mt ‐related genes in the cortices of WT mice after exposure to sevoflurane for 6 h (n = 4–9 per group). F) mRNA expression levels of UPR mt ‐related genes in the cortices of Atf5 ‐cKO mice after exposure to sevoflurane for 6 h (n = 5 per group). G,H) Western blot analysis of cortical samples from Atf5 ‐cKO mice, obtained 24 h after sevoflurane exposure (n = 4–5 per group). I) Left: Mitochondrial function, measured as mitochondrial OCR, determined by assessing respiration of mitochondria isolated from the cortices of mice in control and sevoflurane groups (n = 5 per group). ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. Right: Quantification of OCR after excluding non‐mitochondrial respiration (n = 5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Cell Culture, Immunostaining, Staining, Expressing, Western Blot, Isolation, Control

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane induces mitochondrial dysfunction and fails to induce preconditioning in Gdf15 ‐KO mice. A) Representative TTC‐stained images of MCAO model WT and Gdf15 ‐KO mice after sevoflurane (Sevo) exposure. B) Summary data showing decreased cerebral infarct volume (n = 4–5 per group). C) Volcano plots showing mitochondrial energy metabolism‐related genes whose levels were increased in the cerebral cortex of WT and Gdf15 ‐KO mice 6 h after exposure to sevoflurane (n = 3–5 per group). D) Mitochondrial function, measured as mitochondrial OCR, was determined by assessing the respiration of mitochondria isolated from the cortices of mice in control and sevoflurane groups. ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. E) Quantification of OCR after excluding non‐mitochondrial respiration (n = 3–5 per group). F,G) Western blot analysis of cortical samples obtained from WT and Gdf15 ‐KO mice 24 h after sevoflurane exposure (n = 4–5 per group). H–J) Serum GDF15 protein levels prior to and 3, 6, and 24 h after exposure to sevoflurane in C57BL/6J, WT, and Atf5 ‐cKO mice. K) Comparison of serum GDF15 directly after sevoflurane exposure in C57BL/6J, WT, and Atf5 ‐cKO mice (n = 3–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Staining, Isolation, Control, Western Blot, Comparison

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: ATF5 overexpression in excitatory neurons upregulates GDF15 and enhances mitochondrial function. A) Representative IHC images confirming pAAV‐hSyn‐DIO‐ATF5 expression after stereotaxic injection together with pAAV‐hSyn‐DIO‐EGFP into the cortex of Emx1 cre/+ mice. B) Western blot analysis of cortical samples surrounding the injection site, performed 4 weeks after AAV injection (n = 5 per group). C) Volcano plots showing mitochondrial energy metabolism‐related genes (including Gdf15 ) whose levels were increased around the injection region (n = 5 per group). D) Left: Mitochondrial function, measured as mitochondrial OCR, determined by assessing respiration of mitochondria isolated from the cortex region injected with control or ATF5 virus (n = 5 per group). ADP, oligomycin, CCCP, and antimycin were added sequentially, as indicated by arrows. Right: Quantification of OCR after excluding non‐mitochondrial respiration (n = 5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Over Expression, Expressing, Injection, Western Blot, Isolation, Control, Virus

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: ATF5 overexpression in excitatory neurons provides neuroprotection against ischemic injury. A) Upper panel: Representative image of EGFP (green) and ATF5 (red) expression in the cortex 4 weeks after intravenous injection of pAAV‐hSyn‐DIO‐EGFP. Lower panel: Representative image of CAMKII (green) and ATF5 (red) expression in the cortex 4 weeks after intravenous injection of pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice. B,C) Western blot analysis of ATF5 protein levels in the cortex after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice (n = 5 per group). D) RT‐qPCR analysis of Gdf15 mRNA expression in the cortex after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice (n = 5 per group). E) Serum GDF15 protein levels in Emx1 cre/+ mice 4 weeks after injection with pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 (n = 5 per group). F) Representative TTC‐stained coronal brain sections after intravenous injection of pAAV‐hSyn‐DIO‐EGFP and pAAV‐hSyn‐DIO‐ATF5 in Emx1 cre/+ mice. G) Quantification of cortical infarct volume and neurological score 24 h after MCAO (n = 4–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Over Expression, Expressing, Injection, Western Blot, Quantitative RT-PCR, Staining

Journal: Advanced Science

Article Title: ATF5‐Dependent GDF15 Expression Mediates Anesthesia‐Induced Neuroprotection Against Stroke

doi: 10.1002/advs.202417086

Figure Lengend Snippet: Sevoflurane‐induced activation of ATF5‐dependent UPR mt , GDF15 expression, and mitochondrial protection are absent in cerebral cortices of aged mice. A) Western blot analysis of ATF5 protein levels in the cortex of mice at 2‐, 8‐, 14‐, and 20‐month of age (n = 5 per group). B) Gdf15 mRNA expression in the cortex at 2‐, 8‐, 14‐, and 20‐ months, determined by RT‐qPCR and normalized to β‐actin (n = 3–4 per group). C) Mitochondrial OCR in isolated cortical mitochondria from 2‐, 8‐, 14‐, and 20‐month‐old mice. Left: Representative OCR traces in response to sequential addition of ADP, oligomycin, CCCP, and antimycin. Right: Quantification of basal, state 3, state 4o, and state 3u respiration. D) Volcano plot showing differential expression of 83 mitochondrial energy metabolism‐related genes in the cortex of 20‐month‐old versus 2‐month‐old mice. Red dots, upregulated genes; blue dots, downregulated genes; black dots, non‐significant changes. E) Western blot analysis of proteins in cortical samples from aged mice obtained 24 h after sevoflurane exposure (n = 5 per group). F) Volcano plot showing changes in mitochondrial energy metabolism‐related genes in the cortex of aged mice 6 h after sevoflurane exposure (n = 3–4 per group). G) Left: Mitochondrial function assessed by OCR in isolated cortical mitochondria from aged mice in control and sevoflurane‐treated groups (n = 4–5 per group). Sequential additions of ADP, oligomycin, CCCP, and antimycin are indicated by arrows. Right: Quantification of OCR, excluding non‐mitochondrial respiration (n = 4–5 per group). Values are presented as means ± SD (n.s., not significant; * p < 0.05).

Article Snippet: Conditional Atf5 knockout ( Atf5 cKO) mice were generated by crossing Emx1‐IRES‐Cre knock‐in mice with floxed Atf5 ( Atf5 fl/fl ) mice in which exon 3 was targeted (Cyagen Biosciences, CA, USA).

Techniques: Activation Assay, Expressing, Western Blot, Quantitative RT-PCR, Isolation, Quantitative Proteomics, Control

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of ATF5 protein levels in the intestine, liver, and spleens of Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05 using the Student’s t test). (B) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of Salmonella infection in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 8; ***p < 0.001,****p < 0.0001 using the Student’s t test). (C–E) Oxygen consumption rate (OCR) (C), ATP production (D), and oxidative damage (E) from small intestine samples of Atf5 flox/flox and Atf5 IEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05,**p < 0.001, ***p < 0.001,****p < 0.0001 using the Student’s t test). (F) Mitochondrial membrane potential quantification using TMRE from small intestine samples of Atf5 flox/flox and Atf5 ΔIEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5 ; **** p < 0.0001 using the Student’s t test). (G and H) Changes in bodyweight (G) and feeding (H) of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella . Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05 , ** p < 0.001 using the Student’s t test). (I–K) Colony-forming units (CFU) of intestine (I), liver (J) and spleen (K) samples from Salmonella infected Atf5 flox/flox and Atf5 ΔIEC mice. (n = 5; ns, non-significant, ****p < 0.0001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (n = 5). See for all statistics pertaining to survival analysis. (M and N) Representative histological analysis (M) and pathology score table (N) of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (closed arrowhead represents reduced villi height, open arrowhead represents increased spacing between intestinal crypts, arrow and asterisk represent neutrophilic inflammation and villous fusion, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Western Blot, Control, Infection, Membrane

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-E-cadherin antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (C) Immunoblot analysis and quantification of E-cadherin protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (D) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-ZO-1 antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (E) Immunoblot analysis and quantification of ZO-1 protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; *p < 0.05 using the Student’s t test). (F–H) Immunoblot analysis and quantification of I-FABP (F), DAO (G), and ZO-1 (H) serum levels in Atf5 flox/flox and Atf5 ΔIEC intestinal samples. Coomassie stained SDS-PAGE gels are shown as loading controls (n = 5; ****p < 0.0001 using the Student’s t test). (I and J). Bodyweight (I) and DAI scores (J) of Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05, ****p < 0.0001 using the Student’s t test). (K) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (n = 5). Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (L and M) Representative histological analysis and pathology score table of colon tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Immunohistochemistry, Staining, Western Blot, Control, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of 1.5g/L doxycycline in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ****p < 0.0001 using the Student’s t test). (C and D) Bodyweight of Atf5 flox/flox (C) and Atf5 ΔIEC (D) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, ****p < 0.0001 by the Student’s t test). (E and F) DAI scores of Atf5 flox/flox (E) and Atf5 ΔIEC (F) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , *** p < 0.001 by the Student’s t test). (G and H) Survival of Atf5 flox/flox (G) and Atf5 ΔIEC (H) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control (n = 5). See for all statistics pertaining to survival analysis. (I) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS, in the presence or absence of 1.5 g/L doxycycline pre-treatment. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001, ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (J and K) Representative histological analysis and pathology score table of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline, followed by exposure to 4% DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Western Blot, Control

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Representative image of Atf5 flox/flox and Atf5 ΔIEC mice ( n = 4 ). Scale bar indicates 1.5 cm. (B and C) Bodyweight (B) and feeding behavior (C) of Atf5 flox/flox and Atf5 ΔIEC mice fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 10; *p < 0.05, **p < 0.01 using the Student’s t test). (D) Serum leptin levels in 6-week-old Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E and F) Bodyweight (E) and feeding behavior (F) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin and fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 8; ns, non-significant using the Student’s t test). (G) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < .001 using the Student’s t test). (H and I) Immunoblot analysis and quantification of DAO (H) and I-FABP (I) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (J) Survival of Atf5 flox/flox , Atf5 ΔIEC , and leptin-treated Atf5 ΔIEC mice challenged with Salmonella (n = 5). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Heatmap of differentially expressed genes in intestinal tissues of Atf5 ΔIEC mice compared with Atf5 flox/flox controls, including 269 up and 588 downregulated genes (n = 3). (B) Top 20 URMs predicted based on an ingenuity pathway analysis of genes differentially expressed in intestinal tissues, showing the inferred activation of URMs in Atf5 ΔIEC mice relative to Atf5 flox/flox controls. Z-scores for URM activation are based on observed patterns of gene expression for genes downstream of respective URMs, where the magnitude of the Z score represents evidence for differential activation in Atf5 ΔIEC mice relative Atf5 flox/flox controls, and the sign of the Z score indicates the direction of activation (positive) or repression (negative) in Atf5 ΔIEC mice relative to Atf5 flox/flox controls (n = 3). (C) Top 15 enriched KEGG pathways from differentially expressed genes in Atf5 ΔIEC mice compared with Atf5 flox/flox controls, with distributions of fold change for differentially expressed genes within each KEGG pathway. (D) Cck transcript levels measured by qRT-PCR in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E) Serum CCK levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ***p < 0.001 using the Student’s t test). (F) Serum leptin levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001 using the Student’s t test). (G and H) Bodyweight (G) and feeding behavior (H) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with CCK. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (I) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , * p < 0.05, **p < 0.01 using the Student’s t test). (J and K) Immunoblot analysis and quantification of DAO (J) and I-FABP (K) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Coomassie-stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection, challenged with Salmonella ( n = 5 ). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Activation Assay, Gene Expression, Quantitative RT-PCR, Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A and B) Blood glucose (A) and insulin (B) levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001 using the Student’s t test). (C) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with 2-DG. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (D and E) Immunoblot analysis and quantification of DAO (D) and I-FABP (E) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ****p < 0.0001 using the Student’s t test). (F and G) Clinical scores in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (F) or C . rodentium (G). Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (H and I) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (H) or C . rodentium (I) (n = 5). See for all statistics pertaining to survival analysis. (J–M) CFU counts of liver and spleen samples from Salmonella - (J and K) or C . rodentium -infected (L and M) Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Data represent mean ± standard error of the mean (n = 5; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test).

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Injection, Western Blot, Staining, SDS Page, Infection

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet:

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Recombinant, Virus, Isolation, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Software

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of ATF5 protein levels in the intestine, liver, and spleens of Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05 using the Student’s t test). (B) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of Salmonella infection in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 8; ***p < 0.001,****p < 0.0001 using the Student’s t test). (C–E) Oxygen consumption rate (OCR) (C), ATP production (D), and oxidative damage (E) from small intestine samples of Atf5 flox/flox and Atf5 IEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05,**p < 0.001, ***p < 0.001,****p < 0.0001 using the Student’s t test). (F) Mitochondrial membrane potential quantification using TMRE from small intestine samples of Atf5 flox/flox and Atf5 ΔIEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5 ; **** p < 0.0001 using the Student’s t test). (G and H) Changes in bodyweight (G) and feeding (H) of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella . Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05 , ** p < 0.001 using the Student’s t test). (I–K) Colony-forming units (CFU) of intestine (I), liver (J) and spleen (K) samples from Salmonella infected Atf5 flox/flox and Atf5 ΔIEC mice. (n = 5; ns, non-significant, ****p < 0.0001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (n = 5). See for all statistics pertaining to survival analysis. (M and N) Representative histological analysis (M) and pathology score table (N) of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (closed arrowhead represents reduced villi height, open arrowhead represents increased spacing between intestinal crypts, arrow and asterisk represent neutrophilic inflammation and villous fusion, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Western Blot, Control, Infection, Membrane

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-E-cadherin antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (C) Immunoblot analysis and quantification of E-cadherin protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (D) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-ZO-1 antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (E) Immunoblot analysis and quantification of ZO-1 protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; *p < 0.05 using the Student’s t test). (F–H) Immunoblot analysis and quantification of I-FABP (F), DAO (G), and ZO-1 (H) serum levels in Atf5 flox/flox and Atf5 ΔIEC intestinal samples. Coomassie stained SDS-PAGE gels are shown as loading controls (n = 5; ****p < 0.0001 using the Student’s t test). (I and J). Bodyweight (I) and DAI scores (J) of Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05, ****p < 0.0001 using the Student’s t test). (K) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (n = 5). Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (L and M) Representative histological analysis and pathology score table of colon tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Immunohistochemistry, Staining, Western Blot, Control, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of 1.5g/L doxycycline in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ****p < 0.0001 using the Student’s t test). (C and D) Bodyweight of Atf5 flox/flox (C) and Atf5 ΔIEC (D) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, ****p < 0.0001 by the Student’s t test). (E and F) DAI scores of Atf5 flox/flox (E) and Atf5 ΔIEC (F) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , *** p < 0.001 by the Student’s t test). (G and H) Survival of Atf5 flox/flox (G) and Atf5 ΔIEC (H) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control (n = 5). See for all statistics pertaining to survival analysis. (I) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS, in the presence or absence of 1.5 g/L doxycycline pre-treatment. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001, ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (J and K) Representative histological analysis and pathology score table of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline, followed by exposure to 4% DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Western Blot, Control

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Representative image of Atf5 flox/flox and Atf5 ΔIEC mice ( n = 4 ). Scale bar indicates 1.5 cm. (B and C) Bodyweight (B) and feeding behavior (C) of Atf5 flox/flox and Atf5 ΔIEC mice fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 10; *p < 0.05, **p < 0.01 using the Student’s t test). (D) Serum leptin levels in 6-week-old Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E and F) Bodyweight (E) and feeding behavior (F) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin and fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 8; ns, non-significant using the Student’s t test). (G) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < .001 using the Student’s t test). (H and I) Immunoblot analysis and quantification of DAO (H) and I-FABP (I) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (J) Survival of Atf5 flox/flox , Atf5 ΔIEC , and leptin-treated Atf5 ΔIEC mice challenged with Salmonella (n = 5). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Heatmap of differentially expressed genes in intestinal tissues of Atf5 ΔIEC mice compared with Atf5 flox/flox controls, including 269 up and 588 downregulated genes (n = 3). (B) Top 20 URMs predicted based on an ingenuity pathway analysis of genes differentially expressed in intestinal tissues, showing the inferred activation of URMs in Atf5 ΔIEC mice relative to Atf5 flox/flox controls. Z-scores for URM activation are based on observed patterns of gene expression for genes downstream of respective URMs, where the magnitude of the Z score represents evidence for differential activation in Atf5 ΔIEC mice relative Atf5 flox/flox controls, and the sign of the Z score indicates the direction of activation (positive) or repression (negative) in Atf5 ΔIEC mice relative to Atf5 flox/flox controls (n = 3). (C) Top 15 enriched KEGG pathways from differentially expressed genes in Atf5 ΔIEC mice compared with Atf5 flox/flox controls, with distributions of fold change for differentially expressed genes within each KEGG pathway. (D) Cck transcript levels measured by qRT-PCR in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E) Serum CCK levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ***p < 0.001 using the Student’s t test). (F) Serum leptin levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001 using the Student’s t test). (G and H) Bodyweight (G) and feeding behavior (H) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with CCK. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (I) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , * p < 0.05, **p < 0.01 using the Student’s t test). (J and K) Immunoblot analysis and quantification of DAO (J) and I-FABP (K) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Coomassie-stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection, challenged with Salmonella ( n = 5 ). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Activation Assay, Gene Expression, Quantitative RT-PCR, Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A and B) Blood glucose (A) and insulin (B) levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001 using the Student’s t test). (C) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with 2-DG. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (D and E) Immunoblot analysis and quantification of DAO (D) and I-FABP (E) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ****p < 0.0001 using the Student’s t test). (F and G) Clinical scores in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (F) or C . rodentium (G). Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (H and I) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (H) or C . rodentium (I) (n = 5). See for all statistics pertaining to survival analysis. (J–M) CFU counts of liver and spleen samples from Salmonella - (J and K) or C . rodentium -infected (L and M) Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Data represent mean ± standard error of the mean (n = 5; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test).

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Injection, Western Blot, Staining, SDS Page, Infection

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet:

Article Snippet: Atf5 flox/flox mice, which contain loxP sites flanking exon 3 of the Atf5 gene, were crossed with the B6.Cg-Tg(Vil1-cre)1000Gum/J mouse (Villin Cre ) (The Jackson Laboratory; Bar Harbor, ME) that express the Cre recombinase gene under the control of the villin promoter, generating Atf5 ΔIEC mice (Cyagen Inc.).

Techniques: Recombinant, Virus, Isolation, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Software

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of ATF5 protein levels in the intestine, liver, and spleens of Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05 using the Student’s t test). (B) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of Salmonella infection in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 8; ***p < 0.001,****p < 0.0001 using the Student’s t test). (C–E) Oxygen consumption rate (OCR) (C), ATP production (D), and oxidative damage (E) from small intestine samples of Atf5 flox/flox and Atf5 IEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05,**p < 0.001, ***p < 0.001,****p < 0.0001 using the Student’s t test). (F) Mitochondrial membrane potential quantification using TMRE from small intestine samples of Atf5 flox/flox and Atf5 ΔIEC mice in the presence or absence of Salmonella infection. Data represent mean ± standard error of the mean (n = 5 ; **** p < 0.0001 using the Student’s t test). (G and H) Changes in bodyweight (G) and feeding (H) of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella . Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05 , ** p < 0.001 using the Student’s t test). (I–K) Colony-forming units (CFU) of intestine (I), liver (J) and spleen (K) samples from Salmonella infected Atf5 flox/flox and Atf5 ΔIEC mice. (n = 5; ns, non-significant, ****p < 0.0001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (n = 5). See for all statistics pertaining to survival analysis. (M and N) Representative histological analysis (M) and pathology score table (N) of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice during challenge with Salmonella (closed arrowhead represents reduced villi height, open arrowhead represents increased spacing between intestinal crypts, arrow and asterisk represent neutrophilic inflammation and villous fusion, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Western Blot, Control, Infection, Membrane

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-E-cadherin antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (C) Immunoblot analysis and quantification of E-cadherin protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (D) Immunohistochemistry of Atf5 flox/flox and Atf5 ΔIEC intestinal samples using anti-ZO-1 antibody and DAPI co-stain (n = 3). Representative images shown (boxed region denotes enlarged area). Scale bars indicate 100 μm (wide images) or 10 μm (enlarged images). (E) Immunoblot analysis and quantification of ZO-1 protein levels in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 5; *p < 0.05 using the Student’s t test). (F–H) Immunoblot analysis and quantification of I-FABP (F), DAO (G), and ZO-1 (H) serum levels in Atf5 flox/flox and Atf5 ΔIEC intestinal samples. Coomassie stained SDS-PAGE gels are shown as loading controls (n = 5; ****p < 0.0001 using the Student’s t test). (I and J). Bodyweight (I) and DAI scores (J) of Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5 ; * p < 0.05, ****p < 0.0001 using the Student’s t test). (K) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (n = 5). Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (L and M) Representative histological analysis and pathology score table of colon tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice exposed to DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Immunohistochemistry, Staining, Western Blot, Control, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Immunoblot analysis and quantification of UPR mt -related proteins in the presence or absence of 1.5g/L doxycycline in Atf5 flox/flox and Atf5 ΔIEC mice. Actin was used as a loading control. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (B) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ****p < 0.0001 using the Student’s t test). (C and D) Bodyweight of Atf5 flox/flox (C) and Atf5 ΔIEC (D) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, ****p < 0.0001 by the Student’s t test). (E and F) DAI scores of Atf5 flox/flox (E) and Atf5 ΔIEC (F) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , *** p < 0.001 by the Student’s t test). (G and H) Survival of Atf5 flox/flox (G) and Atf5 ΔIEC (H) mice pre-treated with 1.5 g/L doxycycline and subsequently exposed to 4% DSS or mock control (n = 5). See for all statistics pertaining to survival analysis. (I) Representative image and quantification of colon lengths in Atf5 flox/flox and Atf5 ΔIEC mice exposed to 4% DSS, in the presence or absence of 1.5 g/L doxycycline pre-treatment. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001, ****p < 0.0001 using the Student’s t test). Scale bar, 1 cm. (J and K) Representative histological analysis and pathology score table of small intestine tissue sections from Atf5 flox/flox and Atf5 ΔIEC mice pre-treated with 1.5 g/L doxycycline, followed by exposure to 4% DSS (arrow and asterisk represent epithelial hyperplasia and extensive ulceration and inflammation, respectively; n = 3). Scale bars, 200 μm.

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Western Blot, Control

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Representative image of Atf5 flox/flox and Atf5 ΔIEC mice ( n = 4 ). Scale bar indicates 1.5 cm. (B and C) Bodyweight (B) and feeding behavior (C) of Atf5 flox/flox and Atf5 ΔIEC mice fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 10; *p < 0.05, **p < 0.01 using the Student’s t test). (D) Serum leptin levels in 6-week-old Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E and F) Bodyweight (E) and feeding behavior (F) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin and fed a standard diet. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 8; ns, non-significant using the Student’s t test). (G) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < .001 using the Student’s t test). (H and I) Immunoblot analysis and quantification of DAO (H) and I-FABP (I) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with leptin. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test). (J) Survival of Atf5 flox/flox , Atf5 ΔIEC , and leptin-treated Atf5 ΔIEC mice challenged with Salmonella (n = 5). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A) Heatmap of differentially expressed genes in intestinal tissues of Atf5 ΔIEC mice compared with Atf5 flox/flox controls, including 269 up and 588 downregulated genes (n = 3). (B) Top 20 URMs predicted based on an ingenuity pathway analysis of genes differentially expressed in intestinal tissues, showing the inferred activation of URMs in Atf5 ΔIEC mice relative to Atf5 flox/flox controls. Z-scores for URM activation are based on observed patterns of gene expression for genes downstream of respective URMs, where the magnitude of the Z score represents evidence for differential activation in Atf5 ΔIEC mice relative Atf5 flox/flox controls, and the sign of the Z score indicates the direction of activation (positive) or repression (negative) in Atf5 ΔIEC mice relative to Atf5 flox/flox controls (n = 3). (C) Top 15 enriched KEGG pathways from differentially expressed genes in Atf5 ΔIEC mice compared with Atf5 flox/flox controls, with distributions of fold change for differentially expressed genes within each KEGG pathway. (D) Cck transcript levels measured by qRT-PCR in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ****p < 0.0001 using the Student’s t test). (E) Serum CCK levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; ***p < 0.001 using the Student’s t test). (F) Serum leptin levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant, *p < 0.05, ***p < 0.001 using the Student’s t test). (G and H) Bodyweight (G) and feeding behavior (H) of Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with CCK. Day 0 represents start of experiment at 6 weeks of age. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (I) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Data represent mean ± standard error of the mean (n = 5; ns, non-significant , * p < 0.05, **p < 0.01 using the Student’s t test). (J and K) Immunoblot analysis and quantification of DAO (J) and I-FABP (K) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection. Coomassie-stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; *p < 0.05, **p < 0.01, ***p < 0.001 using the Student’s t test). (L) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal CCK injection, challenged with Salmonella ( n = 5 ). See for all statistics pertaining to survival analysis.

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Activation Assay, Gene Expression, Quantitative RT-PCR, Injection, Western Blot, Staining, SDS Page

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet: (A and B) Blood glucose (A) and insulin (B) levels in Atf5 flox/flox and Atf5 ΔIEC mice. Data represent mean ± standard error of the mean (n = 5; **p < 0.01, ***p < 0.001 using the Student’s t test). (C) Serum FITC-dextran levels in Atf5 flox/flox and Atf5 ΔIEC mice intraperitoneally injected with 2-DG. Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (D and E) Immunoblot analysis and quantification of DAO (D) and I-FABP (E) serum protein levels in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Coomassie stained SDS-PAGE gels are shown as loading controls. Data represent mean ± standard error of the mean (n = 4; **p < 0.01, ****p < 0.0001 using the Student’s t test). (F and G) Clinical scores in Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (F) or C . rodentium (G). Data represent mean ± standard error of the mean (n = 5; ns, non-significant using the Student’s t test). (H and I) Survival of Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection, challenged with Salmonella (H) or C . rodentium (I) (n = 5). See for all statistics pertaining to survival analysis. (J–M) CFU counts of liver and spleen samples from Salmonella - (J and K) or C . rodentium -infected (L and M) Atf5 flox/flox and Atf5 ΔIEC mice, with or without intraperitoneal 2-DG injection. Data represent mean ± standard error of the mean (n = 5; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using the Student’s t test).

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Injection, Western Blot, Staining, SDS Page, Infection

Journal: Cell reports

Article Title: The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response

doi: 10.1016/j.celrep.2022.111789

Figure Lengend Snippet:

Article Snippet: Atf5 flox/flox , Cyagen Inc. , This manuscript.

Techniques: Recombinant, Virus, Isolation, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Software