pex16 antibody  (Proteintech)

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet: The KozakGAL4 knock-in/knock-out strategy, which replaces the coding region of the Pex gene of interest by identifying sgRNA target sites in the 5’ UTR and 3’ UTR. Gray boxes, UTRs; blue boxes, Pex -coding region. (A) Schematics of Pex2 locus and targeting strategy. (B) Schematics of Pex16 locus and targeting strategy. (C) Expression pattern of Pex2 KZ in 3rd instar larval brain. (C’) Repo expression. (C’’) Elav expression. (C’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex2 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain. (D) Expression of Pex16 KZ expression pattern in the third instar Drosophila larval brain. (D’) Repo expression. (D’’) Elav expression. (D’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex16 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain.

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Knock-In, Knock-Out, Expressing

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet:

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Functional Assay, Variant Assay

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet: The KozakGAL4 knock-in/knock-out strategy, which replaces the coding region of the Pex gene of interest by identifying sgRNA target sites in the 5’ UTR and 3’ UTR. Gray boxes, UTRs; blue boxes, Pex -coding region. (A) Schematics of Pex2 locus and targeting strategy. (B) Schematics of Pex16 locus and targeting strategy. (C) Expression pattern of Pex2 KZ in 3rd instar larval brain. (C’) Repo expression. (C’’) Elav expression. (C’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex2 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain. (D) Expression of Pex16 KZ expression pattern in the third instar Drosophila larval brain. (D’) Repo expression. (D’’) Elav expression. (D’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex16 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain.

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Knock-In, Knock-Out, Expressing

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet:

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Functional Assay, Variant Assay

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet: The KozakGAL4 knock-in/knock-out strategy, which replaces the coding region of the Pex gene of interest by identifying sgRNA target sites in the 5’ UTR and 3’ UTR. Gray boxes, UTRs; blue boxes, Pex -coding region. (A) Schematics of Pex2 locus and targeting strategy. (B) Schematics of Pex16 locus and targeting strategy. (C) Expression pattern of Pex2 KZ in 3rd instar larval brain. (C’) Repo expression. (C’’) Elav expression. (C’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex2 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain. (D) Expression of Pex16 KZ expression pattern in the third instar Drosophila larval brain. (D’) Repo expression. (D’’) Elav expression. (D’’’) Merged image with co-localization within the ventral nerve cord, indicating Pex16 KZ expression in both neurons (Elav) and nuclear glia (Elav) in the larval brain.

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Knock-In, Knock-Out, Expressing

Journal: bioRxiv

Article Title: Distinguishing PEX gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in Drosophila

doi: 10.1101/2024.11.14.623590

Figure Lengend Snippet:

Article Snippet: The yw:Pex16 1 line was obtained from Kenji Matsuno, derived from y 1 w 67c23 ; P{w[+mC]y[+mDint2] = EPgy2}Pex16[EY05323]. yw ; Pex16 1 (labeled as Pex16 1 ) For the generation of the human UAS- PEX16 lines for Reference, R176X and del955TCT (F332del), we obtained constructs which were codon-optimized for Drosophila and encoded the human protein from GeneART TM (Thermo Fisher Scientific) (See Supplemental Text ).

Techniques: Functional Assay, Variant Assay

Journal: Nature Communications

Article Title: AAA+ ATPase chaperone p97/VCP FAF2 governs basal pexophagy

doi: 10.1038/s41467-024-53558-x

Figure Lengend Snippet: a Characteristics of the peroxisomal proteins mentioned in this study. b Immunocytochemical images of WT, FAF2-/-, and FAF2-/- HCT116 cells stably expressing 3HA-FAF2 (+3HA-FAF2). Cell nuclei were stained with DAPI. PEX14 and catalase colocalize in WT and FAF2-/- HCT116 cells stably expressing 3HA-FAF2. Peroxisome-deficient cells (i.e., reduced number of catalase-positive dots) were observed in FAF2-/- HCT116 cells. In some FAF2-/- cells, catalase was cytosolic, and PEX14 colocalized with Hsp60 (mitochondrial marker). Higher magnification images of the boxed regions are shown. Scale bars, 10 µm. c . The number of catalase-positive peroxisomes per cell. Peroxisome abundance is significantly reduced in FAF2-/- cells. Dots represent individual data points from three independent experiments. Total number of cells from three independent experiments; n = 53 cells (17, 17, 19 cells/experiments; WT), n = 60 cells (20 cells/experiments; FAF2-/-), and n = 52 cells (17, 17, 18 cells/experiments; FAF2-/- cells + 3HA-FAF2). Bar, median. Statistical significance was calculated using one-way ANOVA; **** p < 0.0001. d Representative images of HA-FAF2 colocalized with peroxisomal catalase. HeLa cells transiently expressing HA-FAF2 were immunostained with anti-HA and anti-catalase antibodies. A line scan (red arrows in the merged panel) shows the colocalization of HA-FAF2 (green line) and catalase (magenta line). Scale bars, 10 µm. e Endogenous FAF2 localized to PMP70-positive peroxisomes. Localization was determined using an in situ proximity ligation assay. WT cells and FAF2-/- cells were reacted with anti-FAF2 and anti-PMP70 antibodies, followed by oligonucleotide-conjugated secondary antibodies (PLA probes). Red dots indicate FAF2-PMP70 interactions. Cell nuclei were stained with DAPI. Scale bars, 10 µm. f , g PMP70, PEX14, PEX16, and catalase are reduced in FAF2-/- cells. WT, FAF2-/-, and FAF2-/- HCT116 cells stably expressing 3HA-FAF2 were immunoblotted with the indicated antibodies. The asterisk indicates an exogenous FAF2 cleavage product that lacks the N-terminal 3HA-tag. h Quantitative analysis of peroxisomal protein levels for cells in f and g . Protein levels were normalized to WT HCT116 cells, which were set to 1. Dots represent individual data points from three independent experiments. Statistical significance was calculated using a one-tailed Welch’s t -test; * p < 0.05, ** p < 0.01, *** p < 0.001; N.S. not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. Source data are provided as a Source Data file.

Article Snippet: The siRNA oligonucleotide sequences are as follows: control, 5’-CGUUAAUCGCGUAUAAUACGCGUAT-3’; USP30, 5’- CCAGAGUCCUGUUCGAUUUtt-3’; OPTN, 5’-GGAGACUGUUGGAAGCGAAtt-3’; NDP52, 5’-CCUUCAUGUGGGUUACUUUtt-3’; TAX1BP1, 5’-CUGAUACACUGGAACACGAtt-3’; FIP200/RB1CC1, 5’- GCCUAGAACAACUAACGAATT-3’; PEX16 5’-GGAUCCUACGGAAGGAGCUUCGGAA-3’; p62, 5’-GAUCUGCGAUGGCUGCAAU-3’; NBR1, 5’-GGAGUGGAUUUACCAGUUA-3’; NSFL1C/p47, 5’-CCAGCAUGUUGUACGGAAA-3’,5’-CCACUUGUUCCCACGAGAA-3’, 5’-UGACUACUUUCCCGAACAA-3’, and 5’-GGAGAGACCAGUAAACCGA-3’; Rep8/UBXD6, 5’-CTGGATGACGAGAATTGGGTA-3’; FAF2, 5’-CTCGTCTGTGGTTGAGTTTAT-3’; ABCD3/PMP70, 5’-GAGUAAGGCUCACUAAAUA-3’, 5’-CCAAAUACCUUGCCACUGU-3’, 5’-CCUCUUAUCUCUCUGGUUA-3’, and 5’-UCAAUAAGCUAGAUACGAA-3’. siRNAs for NPLOC4 and UFD1 knockdown were used as previously reported . siRNAs were transfected into cells using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s protocol.

Techniques: Stable Transfection, Expressing, Staining, Marker, In Situ, Proximity Ligation Assay, One-tailed Test

Journal: Nature Communications

Article Title: AAA+ ATPase chaperone p97/VCP FAF2 governs basal pexophagy

doi: 10.1038/s41467-024-53558-x

Figure Lengend Snippet: a , b HCT116 cells stably expressing 3HA-FAF2 were treated with the chemical crosslinker DSP and then co-immunoprecipitated with anti-HA agarose beads. The samples were immunoblotted with the indicated antibodies. 3HA-FAF2 interacts with PMP70, PEX16, USP30, and p97/VCP, but not PEX26. n = 3 assays. c p97/VCP interacts with FAF2 but not USP30. HeLa cells transiently expressing Flag-tagged p97/VCP WT or the E305Q/E578Q mutant (QQ) were treated with DSP and then co-immunoprecipitated with anti-Flag beads. The samples were immunoblotted with the indicated antibodies. n = 2 assays. d 3HA-FAF2 forms a complex with NPLOC4 and UFD1 but not NSFL1C/p47. HCT116 cells stably expressing 3HA-FAF2 were treated with DSP and then immunoprecipitated with anti-HA agarose beads. The samples were immunoblotted with the indicated antibodies. n = 2 assays. e Schematic diagram of the FAF2 constructs used in this study. UBA, ubiquitin-associated domain; HP, hairpin; CC, coiled-coil domain; UBX, ubiquitin-regulatory X domain. To disrupt interactions with ubiquitin, V47A and L51A substitution were introduced into the UBA domain. deletion; Δ. f FACS-based analysis of the pexophagy flux. Representative FACS data (mKeima-SKL 561/488 ratio) for WT, FAF2-/-, or FAF2-/- cells stably expressing the indicated 3HA-FAF2 mutants with the percentage of pexophagy-positive cells indicated. FAF2 ΔUAS, FAF2 ΔUBA, and FAF2 V47A/L51A restored pexophagy in FAF2-/- cells. The effect of pexophagy suppression by FAF2 ΔHP, ΔCC, and ΔUBX was limited, indicating that the FAF2 UBA and UAS domains are dispensable for pexophagy suppression. deletion; Δ. g Quantitative analysis of the pexophagy flux in f . Dots represent individual data points from three independent experiments. Statistical significance was calculated using one-way ANOVA; ** p < 0.01; **** p < 0.0001; N.S.—not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. deletion; Δ. h Total cell lysates prepared from WT, FAF2-/-, and FAF2-/- HCT116 cells expressing 3HA-FAF2 WT or the indicated mutants were immunoblotted with the indicated antibodies. deletion; Δ. i Quantitative analysis of PMP70 levels in cells from h . PMP70 levels were normalized to WT HCT116 cells, which were set to 1. Dots indicate individual data points from three independent experiments. Statistical significance was calculated using one-way ANOVA; ** p < 0.001; *** p < 0.001; **** p < 0.0001; N.S. not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. deletion; Δ. Source data are provided as a Source Data file.

Article Snippet: The siRNA oligonucleotide sequences are as follows: control, 5’-CGUUAAUCGCGUAUAAUACGCGUAT-3’; USP30, 5’- CCAGAGUCCUGUUCGAUUUtt-3’; OPTN, 5’-GGAGACUGUUGGAAGCGAAtt-3’; NDP52, 5’-CCUUCAUGUGGGUUACUUUtt-3’; TAX1BP1, 5’-CUGAUACACUGGAACACGAtt-3’; FIP200/RB1CC1, 5’- GCCUAGAACAACUAACGAATT-3’; PEX16 5’-GGAUCCUACGGAAGGAGCUUCGGAA-3’; p62, 5’-GAUCUGCGAUGGCUGCAAU-3’; NBR1, 5’-GGAGUGGAUUUACCAGUUA-3’; NSFL1C/p47, 5’-CCAGCAUGUUGUACGGAAA-3’,5’-CCACUUGUUCCCACGAGAA-3’, 5’-UGACUACUUUCCCGAACAA-3’, and 5’-GGAGAGACCAGUAAACCGA-3’; Rep8/UBXD6, 5’-CTGGATGACGAGAATTGGGTA-3’; FAF2, 5’-CTCGTCTGTGGTTGAGTTTAT-3’; ABCD3/PMP70, 5’-GAGUAAGGCUCACUAAAUA-3’, 5’-CCAAAUACCUUGCCACUGU-3’, 5’-CCUCUUAUCUCUCUGGUUA-3’, and 5’-UCAAUAAGCUAGAUACGAA-3’. siRNAs for NPLOC4 and UFD1 knockdown were used as previously reported . siRNAs were transfected into cells using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s protocol.

Techniques: Stable Transfection, Expressing, Immunoprecipitation, Mutagenesis, Construct

Journal: Nature Communications

Article Title: AAA+ ATPase chaperone p97/VCP FAF2 governs basal pexophagy

doi: 10.1038/s41467-024-53558-x

Figure Lengend Snippet: a Characteristics of the peroxisomal proteins mentioned in this study. b Immunocytochemical images of WT, FAF2-/-, and FAF2-/- HCT116 cells stably expressing 3HA-FAF2 (+3HA-FAF2). Cell nuclei were stained with DAPI. PEX14 and catalase colocalize in WT and FAF2-/- HCT116 cells stably expressing 3HA-FAF2. Peroxisome-deficient cells (i.e., reduced number of catalase-positive dots) were observed in FAF2-/- HCT116 cells. In some FAF2-/- cells, catalase was cytosolic, and PEX14 colocalized with Hsp60 (mitochondrial marker). Higher magnification images of the boxed regions are shown. Scale bars, 10 µm. c . The number of catalase-positive peroxisomes per cell. Peroxisome abundance is significantly reduced in FAF2-/- cells. Dots represent individual data points from three independent experiments. Total number of cells from three independent experiments; n = 53 cells (17, 17, 19 cells/experiments; WT), n = 60 cells (20 cells/experiments; FAF2-/-), and n = 52 cells (17, 17, 18 cells/experiments; FAF2-/- cells + 3HA-FAF2). Bar, median. Statistical significance was calculated using one-way ANOVA; **** p < 0.0001. d Representative images of HA-FAF2 colocalized with peroxisomal catalase. HeLa cells transiently expressing HA-FAF2 were immunostained with anti-HA and anti-catalase antibodies. A line scan (red arrows in the merged panel) shows the colocalization of HA-FAF2 (green line) and catalase (magenta line). Scale bars, 10 µm. e Endogenous FAF2 localized to PMP70-positive peroxisomes. Localization was determined using an in situ proximity ligation assay. WT cells and FAF2-/- cells were reacted with anti-FAF2 and anti-PMP70 antibodies, followed by oligonucleotide-conjugated secondary antibodies (PLA probes). Red dots indicate FAF2-PMP70 interactions. Cell nuclei were stained with DAPI. Scale bars, 10 µm. f , g PMP70, PEX14, PEX16, and catalase are reduced in FAF2-/- cells. WT, FAF2-/-, and FAF2-/- HCT116 cells stably expressing 3HA-FAF2 were immunoblotted with the indicated antibodies. The asterisk indicates an exogenous FAF2 cleavage product that lacks the N-terminal 3HA-tag. h Quantitative analysis of peroxisomal protein levels for cells in f and g . Protein levels were normalized to WT HCT116 cells, which were set to 1. Dots represent individual data points from three independent experiments. Statistical significance was calculated using a one-tailed Welch’s t -test; * p < 0.05, ** p < 0.01, *** p < 0.001; N.S. not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. Source data are provided as a Source Data file.

Article Snippet: For siRNA knockdown of USP30, OPTN, NDP52, TAX1BP1, FIP200/RB1CC1, and PEX16, Silencer Select Pre-designed siRNAs (IDs s39400, s19719, s19996, s16986, and s18994) and Stealth siRNA (HSS145149) were purchased from Thermo Fisher Scientific. siGENOME siRNA (D-010230-02-0005, D-010522-01-0005, and M-009909-01-0005) and custom siRNAs (LP_23815) for knockdown of p62, NBR1, NSFL1C/p47, and PMP70 were purchased from Dharmacon.

Techniques: Stable Transfection, Expressing, Staining, Marker, In Situ, Proximity Ligation Assay, One-tailed Test

Journal: Nature Communications

Article Title: AAA+ ATPase chaperone p97/VCP FAF2 governs basal pexophagy

doi: 10.1038/s41467-024-53558-x

Figure Lengend Snippet: a , b HCT116 cells stably expressing 3HA-FAF2 were treated with the chemical crosslinker DSP and then co-immunoprecipitated with anti-HA agarose beads. The samples were immunoblotted with the indicated antibodies. 3HA-FAF2 interacts with PMP70, PEX16, USP30, and p97/VCP, but not PEX26. n = 3 assays. c p97/VCP interacts with FAF2 but not USP30. HeLa cells transiently expressing Flag-tagged p97/VCP WT or the E305Q/E578Q mutant (QQ) were treated with DSP and then co-immunoprecipitated with anti-Flag beads. The samples were immunoblotted with the indicated antibodies. n = 2 assays. d 3HA-FAF2 forms a complex with NPLOC4 and UFD1 but not NSFL1C/p47. HCT116 cells stably expressing 3HA-FAF2 were treated with DSP and then immunoprecipitated with anti-HA agarose beads. The samples were immunoblotted with the indicated antibodies. n = 2 assays. e Schematic diagram of the FAF2 constructs used in this study. UBA, ubiquitin-associated domain; HP, hairpin; CC, coiled-coil domain; UBX, ubiquitin-regulatory X domain. To disrupt interactions with ubiquitin, V47A and L51A substitution were introduced into the UBA domain. deletion; Δ. f FACS-based analysis of the pexophagy flux. Representative FACS data (mKeima-SKL 561/488 ratio) for WT, FAF2-/-, or FAF2-/- cells stably expressing the indicated 3HA-FAF2 mutants with the percentage of pexophagy-positive cells indicated. FAF2 ΔUAS, FAF2 ΔUBA, and FAF2 V47A/L51A restored pexophagy in FAF2-/- cells. The effect of pexophagy suppression by FAF2 ΔHP, ΔCC, and ΔUBX was limited, indicating that the FAF2 UBA and UAS domains are dispensable for pexophagy suppression. deletion; Δ. g Quantitative analysis of the pexophagy flux in f . Dots represent individual data points from three independent experiments. Statistical significance was calculated using one-way ANOVA; ** p < 0.01; **** p < 0.0001; N.S.—not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. deletion; Δ. h Total cell lysates prepared from WT, FAF2-/-, and FAF2-/- HCT116 cells expressing 3HA-FAF2 WT or the indicated mutants were immunoblotted with the indicated antibodies. deletion; Δ. i Quantitative analysis of PMP70 levels in cells from h . PMP70 levels were normalized to WT HCT116 cells, which were set to 1. Dots indicate individual data points from three independent experiments. Statistical significance was calculated using one-way ANOVA; ** p < 0.001; *** p < 0.001; **** p < 0.0001; N.S. not significant. The center lines correspond to the medians, and the box limits indicate the 25th and 75th percentiles. The box-plot whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. deletion; Δ. Source data are provided as a Source Data file.

Article Snippet: For siRNA knockdown of USP30, OPTN, NDP52, TAX1BP1, FIP200/RB1CC1, and PEX16, Silencer Select Pre-designed siRNAs (IDs s39400, s19719, s19996, s16986, and s18994) and Stealth siRNA (HSS145149) were purchased from Thermo Fisher Scientific. siGENOME siRNA (D-010230-02-0005, D-010522-01-0005, and M-009909-01-0005) and custom siRNAs (LP_23815) for knockdown of p62, NBR1, NSFL1C/p47, and PMP70 were purchased from Dharmacon.

Techniques: Stable Transfection, Expressing, Immunoprecipitation, Mutagenesis, Construct

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Acid Assay, Cholesterol Assay, Colorimetric Assay

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Acid Assay, Cholesterol Assay, Colorimetric Assay

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: Mice and Treatment: Pex16 -floxed mice ( Pex16 fl/fl mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #034155) were crossed with transgenic mice expressing Albumin-Cre recombinase (Alb-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #003574) and/or expressing adiponectin-Cre recombinase (adipoQ-Cre mice; purchased from Jackson Laboratory, Bar Harbour, ME, USA, strain number #028020) to create the Pex16 Alb-Cre mice and adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice, respectively.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Acid Assay, Cholesterol Assay, Bioassay, Colorimetric Assay, Control

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Acid Assay, Cholesterol Assay, Bioassay, Colorimetric Assay, Control

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing