Journal: Scientific Reports

Article Title: Influenza virus infection affects insulin signaling, fatty acid-metabolizing enzyme expressions, and the tricarboxylic acid cycle in mice

doi: 10.1038/s41598-020-67879-6

Figure Lengend Snippet: Expressions of energy metabolism-related genes in the liver. Mice were intranasally inoculated with PBS alone or PBS comprising PR8 virus, and liver samples were collected at 1, 3, and 6 dpi. The gene expressions of Pck2 , Cd36 , Pnpla2 , Acox1 , and Cpt1b were normalized with that of 18S from real-time PCR analyses. Gene expressions of the PR8 virus-infected mice are presented as fold changes relative to those of the control mice at each time point. Bars represent means ± SEM of 3 or 4 animals. Gray and black bars indicate data from control and PR8 virus-infected mice, respectively; * p < 0.05, unpaired t test using the Holm–Sidak correction method with an alpha value of 0.05, control vs. PR8 virus-infected mice at each time point.

Article Snippet: The gene expressions of Cluster of differentiation 36 ( Cd36 , Mm00432403_m1), Patatin Like Phospholipase Domain Containing 2 ( Pnpla2 , Mm00503040_m1), Acyl-CoA Oxidase 1 ( Acox1 , Mm01246834_m1), Carnitine Palmitoyltransferase 1 Beta ( Cpt1b , Mm00487191_g1), and Phosphoenolpyruvate Carboxykinase 2 ( Pck2 , Mm00551411_m1) were quantified using real-time PCR with a StepOne Real-Time PCR system (Applied Biosystems, Foster City, CA) with TaqMan probes (Applied Biosystems).

Techniques: Virus, Real-time Polymerase Chain Reaction, Infection, Control

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: KEGG pathway analysis of differentially expressed genes associated with NSCLC

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Starch

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: The expression levels of mRNA COL12A1 (G) , COL1A2 (D) , COL4A1 (E) , SPARC (K) , COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A) , ACOX1 (B) , CPT2 (H) , SQRDL (L) , HMGCS2 (J) , ETHE1 (I) and TST (M) were decreased. * P < 0.05.

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Expressing

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: The expression levels of protein COL12A1 (G) , COL1A2 (D) , COL4A1 (E) , SPARC (K) , COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A) , ACOX1 (B) , CPT2 (H) , SQRDL (L) , HMGCS2 (J) , ETHE1 (I) and TST (M) were decreased. * P < 0.05.

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Expressing

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: Primer pairs of different genes

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques:

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Changes in lipid metabolism in ft/ft mouse epidermis. ( a ) Microarray analysis showing a subset of genes involved in FA metabolism in mouse epidermis. The full gene list and respective fold changes are provided in Table 3 . ( b ) Relative mRNA level of Acox1 and Hsd17b4 in the epidermis of mice (n = 9–10). ( c, d ) Protein abundance of ACOX1 in mouse epidermis. The dashed line indicates the dermal‒epidermal boundary. Bar = 50 μm ( e ) ACOX activity measured in mouse epidermal cells (expressed as mU per μg of proteins, n = 5–7). Relative mRNA expression of ( f ) Acot5 , Acot8 , ( g ) Crot , and ( h ) PPAR mRNA, Ppar , isoforms in epidermal samples from CTRL and ft/ft mice (n = 9–10). Data were analyzed with a Student’s t -test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. CTRL, control; FA, fatty acid; H 2 O 2 , hydrogen peroxide; PPAR, peroxisome proliferator–activated receptor.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Microarray, Activity Assay, Expressing

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: PCR Array Gene List and Respective Fold Changes

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Sequencing, Binding Assay, Transferring

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Detection of ACOX1 in human AD skin and overexpression of ACOX1 in HEEs. ( a, b ) Representative immunostaining showing the protein abundance of ACOX1 in ADL) and in ADNL epidermis compared with the epidermis of healthy donors (CTRL). The dashed line indicates the dermal‒epidermal boundary. Bar = 30 μm. ( c ) Representative H&E staining of HEEs generated with KCs infected with lentivirus containing either pHR-SFFV-ACOX1 (ACOX1 OE) or pHR-SFFV-Puro CTRL(Puro) vector. Bar = 50 μm. ( d ) mRNA and ( e, f ) protein levels of ACOX1 in HEEs overexpressing ACOX1 compared with those in their Puro CTRLs (n = 8). The dashed line indicates the basal epidermal layer. Bar = 50 μm. Data were analyzed with a paired Student’s t -test. ∗∗∗ P < 0.001. AD, atopic dermatitis; ADL, lesional atopic dermatitis; ADNL, nonlesional atopic dermatitis; CTRL, control; HEE, human epidermal equivalent; KC, keratinocyte.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Over Expression, Immunostaining, Staining, Generated, Infection, Plasmid Preparation

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Analysis of HEEs overexpressing ACOX1. ( a ) Ultrastructural analysis showing LB secretion (upper panel, arrows), LB numbers (lower panel, arrows), and morphology (insets) in HEEs overexpressing ACOX1 (right panel) and in their Puro controls (left panel). Osmium tetroxide after fixation. Bar = 250 nm or 125 nm (inset). ( b ) LB numbers and ( c ) quantified secretion areas in HEEs in eight randomly selected fields per group (n = 2). ( d ) TEER and ( e ) LY penetration assay (green) in HEEs. Nuclei were counterstained with DAPI (blue). Bar = 50 μm. (n = 3). ( f ) Representative Ki-67 staining and the number of Ki-67‒positive nuclei in HEEs overexpressing ACOX1 compared with those in their Puro controls (n = 5). ( g ) Heat map showing the fold changes in the mRNA level of inflammation-related genes in HEEs (n = 5–7). Data were analyzed with a paired Student’s t -test. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. HEE, human epidermal equivalent; LB, lamellar body; LY, Lucifer yellow; SC, stratum corneum; SG, stratum granulosum; TEER, transepithelial electrical resistance.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Staining

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Lipidomic analysis of HEEs overexpressing ACOX1. ( a ) d18:1 Cers(NS) (n = 3) and ( b ) FFA (n = 3) species in HEEs generated with KCs infected with lentivirus containing either pHR-SFFV-ACOX1 (ACOX1 OE) or pHR-SFFV-Puro control (Puro) vector. Data are shown as analyte/IS ratio (AU) per mg protein or as the relative percentage of total lipid species. Data were analyzed with a Student’s t -test. ∗ P < 0.05. AU: arbitrary unit; Cer, ceramide; FFA, free fatty acid; HEE, human epidermal equivalent; IS, internal standard; KC, keratinocyte; MUFA, monounsaturated fatty acid; SFA, saturated fatty acid.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Generated, Infection, Plasmid Preparation

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Fatty acid synthesis and elongation in ft/ft mouse epidermis. ( a ) Relative mRNA levels of Fasn , Elov11 , Elov4 , and Elov6 in the epidermis of ft/ft mice compared with those in the epidermis of the CTRL mice (n = 9–10). ( b ) Representative western blot and immunostaining showing protein abundance of ELOVL1 in the epidermis of CTRL and ft/ft mice. ( c ) qPCR showing the relative mRNA level of Acox1 (left panel) and its protein abundance (right panel) in the epidermis of ft/ft mice uncovered (CTRL) or covered with an occlusive dressing to reduce TEWL (n = 5). The dashed line indicates the dermal‒epidermal boundary. Bar = 50 μm. Data were analyzed with a Student’s t -test. ∗ P < 0.05 and ∗∗∗ P < 0.001. CTRL, control; TEWL, transepidermal water loss.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Western Blot, Immunostaining

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Metabolic and ultrastructural analysis of Flg -KO mouse epidermis. ( a ) Microarray analysis showing fold changes of a subset of genes involved in FA metabolism in the epidermis of Flg -KO mice compared with those in the epidermis of CTRLs. ( b ) Relative Acox1 mRNA level (n = 10) and protein abundance as well as ( c ) ACOX activity in CTRL and Flg -KO mouse epidermis (n = 5). ( d ) Relative levels of PPAR mRNA, Ppar , isoforms in mouse epidermis (n = 10). ( e ) Ultrastructural analysis of Flg -KO mouse epidermis showing LB morphology and secretion (arrows). Osmium tetroxide after fixation. Bar = 250 nm. ( f ) Relative mRNA level of Glut1 in mouse epidermis (n = 5). ( g ) Glucose consumption and ( d ) lactate production by epidermal sheets of CTRL and Flg -KO mice (n = 5). ( h ) Intracellular levels of metabolites and Krebs cycle intermediates quantified by LC‒MS in the epidermis of Flg -KO mice compared with those in the epidermis of CTRLs. Fold changes between the mean values of five mice per group are shown (n = 5). Data were analyzed with a Student’s t -test. The list of genes featured on the array and of all quantified metabolites as well as respective fold changes are provided in Tables 3 and . CTRL, control; Flg -KO, Flg -knockout; KO, knockout; LB, lamellar body; LC‒MS, liquid chromatography‒mass spectrometry; PPAR, peroxisome proliferator–activated receptor; SC, stratum corneum; SG, stratum granulosum.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Microarray, Activity Assay, Knock-Out

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: MC903 mouse model of ADL ( a ) Representative H&E staining of ear sections (bar = 200 μm) of MC903-treated mice compared with that of the vehicle-treated CTRLS (n = 7) and ( b ) TEWL values measured on the ears of mice (n = 7). ( c ) qPCR and immunostaining showing mRNA and protein level of ACOX1 in the epidermis of mice treated with MC903 or vehicle. The dashed line indicates the dermal‒epidermal boundary. Bar = 50 μm. mRNA level of ( d ) Acot5 ; ( e ) Crot ; ( f ) PPAR mRNA, Ppar , isoforms; and Fabp5 in the epidermis of mice treated with MC903 or vehicle (n = 7). ( g ) mRNA level and protein abundance of GLUT1 and ( h ) mRNA level of key enzymes of glycolysis as well as of ( i ) Ldha and Pdk1 in the epidermis of mice treated with MC903 or vehicle (n = 7). The dashed line indicates the dermal‒epidermal boundary. Bar = 50 μm. Data were analyzed with a Student’s t -test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. ADL, lesional atopic dermatitis; CTRL, control; PPAR, peroxisome proliferator–activated receptor; TEWL, transepidermal water loss.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Staining, Immunostaining

Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: IMQ mouse model and biopsies from patients with PSO. ( a ) Representative H&E staining of ear sections (bar = 200 μm) of mice treated with 5% IMQ cream or with petroleum jelly (CTRL) (n = 7) and ( b ) TEWL values measured on the ears of mice (n = 7). ( c ) qPCR and immunostaining showing mRNA and protein level of ACOX1 in the epidermis of mice treated with 5% IMQ or with petroleum jelly. The dashed line indicates the dermal‒epidermal boundary. Bar = 50 μm. ( d ) Immunostaining of ACOX1 in the epidermis of patients with PSO and healthy donors (CTRL). The dashed line indicates the dermal‒epidermal boundary. Bar = 30 μm. Data were analyzed with a Student’s t -test. ∗∗∗ P < 0.001. CTRL, control; IMQ, imiquimod; PSO, psoriasis; TEWL, transepidermal water loss.

Article Snippet: The lentiviral vector containing human ACOX1 (NM_004035) was generated by cloning human ACOX1 cDNA from the pCMV-ACOX1 (OriGene Technologies, Rockville, MD) plasmid into the BamHI/NotI site of the lentiviral pHR-SIN-CSGW vector (kindly provided by Mary Collins, University College London, London, United Kingdom), thereby generating pHR-SFFV-hACOX1.

Techniques: Staining, Immunostaining