endogenous apmap (Nikon)

Nikon endogenous apmap

Figure 1. <t>APMAP</t> localizes to the ER in mammalian cells and Drosophila FB (A) APMAP domains. APMAPFL (full-length human APMAP, transmembrane [TM] domain, and 6-bladed β-propeller region). Paraoxonases (PON1 and PON2, 6-bladed β-propeller domains).

Endogenous Apmap, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 1. APMAP localizes to the ER in mammalian cells and Drosophila FB (A) APMAP domains. APMAPFL (full-length human APMAP, transmembrane [TM] domain, and 6-bladed β-propeller region). Paraoxonases (PON1 and PON2, 6-bladed β-propeller domains).

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 1. APMAP localizes to the ER in mammalian cells and Drosophila FB (A) APMAP domains. APMAPFL (full-length human APMAP, transmembrane [TM] domain, and 6-bladed β-propeller region). Paraoxonases (PON1 and PON2, 6-bladed β-propeller domains).

Article Snippet: The secondary antibodies are donkey anti-mouse AF488 (Thermo Fisher Scientific; A21202), donkey anti-mouse AF594 (Thermo Fisher Scientific; A21203), donkey anti-rabbit AF488 (Thermo Fisher Scientific; A21206), donkey anti-rabbit AF594 (Thermo Fisher Scientific; A21207), donkey anti-goat AF488 (Thermo Fisher Scientific; A11055) and donkey anti-goat AF594 (Thermo Fisher Scientific; A11058) used at a dilution of 1:400 and incubated for 30 min. LDs were visualized by staining the cells with MDH (1:1,000; Abgent; SM1000a) for 15 min. Confocal fluorescence microscopy and Image analysis Confocal microscopy of the cells was performed using the following equipment according to the experiment – a) 63x oil-immersion objective lens in a Zeiss laser scanning 780 confocal and Zeiss LSM880 Airyscan microscope; b) Live-cell imaging of endogenous APMAP using 4-100x objectives in Nikon TiE inverted microscope with a Yokogawa CSU-W1 spinning disk+SoRa module.

Techniques:

Figure 2. APMAP is a type II integral membrane protein (A) Protease protection assay. Huh7 cells over-expressing EGFP-KDEL, APMAPFL-EGFP, and Reticulon4a-EGFP treated with digitonin for selective per- meabilization, followed by PK for proteolysis of GFP tag. (B) Live-cell confocal of Huh7 cells expressing EGFP-KDEL, APMAPFL-EGFP, and Reticulon4a-EGFP with 0.01% digitonin and 50 μg/mL PK. Scale bar, 10 μm. (C) Cartoon of APMAP type II membrane topology. (D) Volcano plot of TurboID-based proteomics showing APMAP over-expressed Huh7 cells compared with control (untransfected) cells. Log2 fold enrichment (x axis), −log10 of p values (y axis). (E) Proteins-of-interest from APMAP TurboID proteomics.

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 2. APMAP is a type II integral membrane protein (A) Protease protection assay. Huh7 cells over-expressing EGFP-KDEL, APMAPFL-EGFP, and Reticulon4a-EGFP treated with digitonin for selective per- meabilization, followed by PK for proteolysis of GFP tag. (B) Live-cell confocal of Huh7 cells expressing EGFP-KDEL, APMAPFL-EGFP, and Reticulon4a-EGFP with 0.01% digitonin and 50 μg/mL PK. Scale bar, 10 μm. (C) Cartoon of APMAP type II membrane topology. (D) Volcano plot of TurboID-based proteomics showing APMAP over-expressed Huh7 cells compared with control (untransfected) cells. Log2 fold enrichment (x axis), −log10 of p values (y axis). (E) Proteins-of-interest from APMAP TurboID proteomics.

Techniques: Membrane, Expressing, Control

Figure 3. APMAP depletion causes perturbed ER morphology and LD accumulation (A) (Top) APMAP mRNA levels, (bottom) APMAP protein (Western) (N = 3; ****p < 0.0001; ordinary one-way ANOVA). (B) Airyscan confocal of Huh7 cells with scrambled siRNA (siCtrl) or siAPMAP. IF staining α-calnexin (green) or α-CLIMP63 (magenta). Scale bar, 5 μm.

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 3. APMAP depletion causes perturbed ER morphology and LD accumulation (A) (Top) APMAP mRNA levels, (bottom) APMAP protein (Western) (N = 3; ****p < 0.0001; ordinary one-way ANOVA). (B) Airyscan confocal of Huh7 cells with scrambled siRNA (siCtrl) or siAPMAP. IF staining α-calnexin (green) or α-CLIMP63 (magenta). Scale bar, 5 μm.

Techniques: Western Blot, Staining

Figure 4. Drosophila and zebrafish depletion of APMAP alters TG storage and lipoproteins (A) Confocal of larval fat-body (FB) from control (Dcg-Gal4 line) and dAPMAP-RNAi(FB-specific). LDs with MDH (magenta). Scale bar, 10 μm. (B) Relative whole-larval TG from control (Dcg-Gal4) and dAPMAP-RNAi(FB-specific) (N = 3; **p < 0.0021; two-tailed unpaired t test).

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 4. Drosophila and zebrafish depletion of APMAP alters TG storage and lipoproteins (A) Confocal of larval fat-body (FB) from control (Dcg-Gal4 line) and dAPMAP-RNAi(FB-specific). LDs with MDH (magenta). Scale bar, 10 μm. (B) Relative whole-larval TG from control (Dcg-Gal4) and dAPMAP-RNAi(FB-specific) (N = 3; **p < 0.0021; two-tailed unpaired t test).

Techniques: Control, Two Tailed Test

Figure 5. APMAP loss alters cellular redox homeostasis, which is rescued by chemical antioxidant NAC (A) Oxidized BODIPY-C11 in Huh7 cells NAC: N-acetyl cysteine (NAC). Each dot represents oxidized BODIPY-C11 of a cell depicted as ratio of green(oxidized) over red(reduced) (N = 3; n > 50; ****p < 0.0001, **p < 0.0021, *p < 0.0332, ordinary one-way ANOVA with Sidak’s multiple comparison α = 0.05).

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 5. APMAP loss alters cellular redox homeostasis, which is rescued by chemical antioxidant NAC (A) Oxidized BODIPY-C11 in Huh7 cells NAC: N-acetyl cysteine (NAC). Each dot represents oxidized BODIPY-C11 of a cell depicted as ratio of green(oxidized) over red(reduced) (N = 3; n > 50; ****p < 0.0001, **p < 0.0021, *p < 0.0332, ordinary one-way ANOVA with Sidak’s multiple comparison α = 0.05).

Techniques: Comparison

Figure 6. APMAP depletion alters lipid saturation and increases ceramides (A) Volcano plot depicting total abundance of lipid species in Huh7 cells, determined through LC-MS/MS. Each data point represents average level of individual fatty acid of a lipid species. Log2 fold-enrichment of fatty acid (x axis) plotted against −log10 p values from unpaired t test (y axis).

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 6. APMAP depletion alters lipid saturation and increases ceramides (A) Volcano plot depicting total abundance of lipid species in Huh7 cells, determined through LC-MS/MS. Each data point represents average level of individual fatty acid of a lipid species. Log2 fold-enrichment of fatty acid (x axis) plotted against −log10 p values from unpaired t test (y axis).

Techniques: Liquid Chromatography with Mass Spectroscopy

Figure 7. Suppression of ceramide synthesis and NAC treatment in APMAP-depleted cells rescues intracellular ApoB accumulation (A) Schematic of ceramide synthesis with ER stress and signaling. Myriocin (Myr or M) and fumonisin B (Fum or F) are inhibitors of serine palmitoyltransferase (SPT) and CerS, respectively. Representative IF images of ceramide in WT and fumonisin B-treated Huh7 cells.

Journal: Developmental cell

Article Title: Paraoxonase-like APMAP maintains endoplasmic-reticulum-associated lipid and lipoprotein homeostasis.

doi: 10.1016/j.devcel.2025.04.008

Figure Lengend Snippet: Figure 7. Suppression of ceramide synthesis and NAC treatment in APMAP-depleted cells rescues intracellular ApoB accumulation (A) Schematic of ceramide synthesis with ER stress and signaling. Myriocin (Myr or M) and fumonisin B (Fum or F) are inhibitors of serine palmitoyltransferase (SPT) and CerS, respectively. Representative IF images of ceramide in WT and fumonisin B-treated Huh7 cells.

Techniques:

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