Journal: Nature Communications

Article Title: The NERP-4–SNAT2 axis regulates pancreatic β-cell maintenance and function

doi: 10.1038/s41467-023-43976-8

Figure Lengend Snippet: a Volcano plot depicting a comparison of proteins captured by NERP-4 or NERP-2. Data are shown at the protein level and are annotated using the UniProt mouse database. Y -axis = −Log10 (adjusted P value), X -axis = Log2 fold change compared to the other samples ( n = 3 independent experiments). b Target candidates identified by the TriCEPS™-based ligand–receptor capture method. SCRB2: lysosome membrane protein 2. c Binding of [ 125 I]-Y-NERP-4[8–19] to the membrane of SNAT2-OE HEK293 cells with or without unlabelled NERP-4 ( n = 4 biological replicates). d Binding of [ 125 I]-Y-NERP-4[8–19] with or without unlabelled NERP-4 in SNAT2-overexpressing (OE) HEK293 cells ( n = 4 biological replicates). e mRNA levels of Snat1 , Snat2 , Snat3 , Snat4 , and Snat5 in C57BL/6 J mouse islets and MIN6-K8 cells ( n = 5 biological replicates). f Binding of [ 125 I]-Y-NERP-4[8–19] to mock, SNAT2-, SNAT3-, SNAT4-, or SNAT5-OE HEK293 cells ( n = 3 biological replicates). Representative results of two independent experiments ( c – f ). Data are mean ± s.e.m ( c – f ). Differential protein abundance was tested using a statistical ANOVA model followed by multiple testing corrections ( a , b ). One-way ANOVA and Fisher’s LSD test, #P value, SNAT2 OE vs. SNAT2 OE plus unlabelled NERP-4; *P value, SNAT2 OE vs. mock ( c ). One-way ANOVA and Tukey’s multiple comparisons test ( d , e ). Two-way ANOVA and Tukey’s multiple comparisons test, * P = 0.0444, ** P = 0.0051, *** P < 0.0001 vs. mock; not significant, SNAT3/SNAT4/SNAT5 vs. mock ( f ). Source data are provided as a file.

Article Snippet: Human SNAT2/SLC38A2 (Myc-DDK–tagged)-CMV 6-entry vector was purchased from Origene (RC201892).

Techniques: Comparison, Membrane, Binding Assay

Journal: Nature Communications

Article Title: The NERP-4–SNAT2 axis regulates pancreatic β-cell maintenance and function

doi: 10.1038/s41467-023-43976-8

Figure Lengend Snippet: NERP-4–induced [ 14 C]- l -glutamine uptake into human islets ( a , n = 4 biological replicates), C57BL/6 J mouse islets ( b , n = 6, 5 biological replicates), and MIN6-K8 cells ( c , n = 4 biological replicates). d [ 14 C]- l -Alanine uptake into MIN6-K8 cells ( n = 4 biological replicates). [ 14 C]- l -Glutamine ( e ) and [ 14 C]- l -alanine ( f ) uptake in si Snat2 -MIN6-K8 cells ( n = 4 biological replicates). [ 14 C]- l -Glutamine ( g ) and [ 14 C]- l -alanine ( h ) uptake by MIN6-K8 cells with or without NERP-4 and MeAIB ( n = 4 biological replicates). i Representative Fura-2-AM ratios in si Snat2 –MIN6-K8 cells in response to NERP-4 ( n = 8 cells), and average iAUC (9–20 min) of [Ca 2+ ] i ( n = 8 cells). Effects of Snat2 knockdown ( j , n = 6 biological replicates) and MeAIB ( k , n = 8 biological replicates) on NERP-4–induced GSIS in MIN6-K8 cells. Effect of MeAIB on NERP-4–induced GSIS in human islets ( l , n = 5 biological replicates) and C57BL/6 J mouse islets ( m , n = 6 biological replicates). n Concentration dependence of MeAIB uptake into MIN6-K8 cells in the presence or absence of NERP-4 ( n = 4 biological replicates). Inset shows 1 or 10 μM MeAIB uptake. All experiments were performed under 16.7 mM glucose. Representative results of two independent experiments ( b – n ). Data are mean ± s.e.m ( a – n ). One-way ANOVA and Tukey’s multiple comparisons test ( a , c , e – h , j – n ). Unpaired two-tailed Student’s t -test ( b , d , i ). Source data are provided as a file.

Article Snippet: Human SNAT2/SLC38A2 (Myc-DDK–tagged)-CMV 6-entry vector was purchased from Origene (RC201892).

Techniques: Concentration Assay, Two Tailed Test

Journal: Nature Communications

Article Title: The NERP-4–SNAT2 axis regulates pancreatic β-cell maintenance and function

doi: 10.1038/s41467-023-43976-8

Figure Lengend Snippet: a, Snat2 mRNA amounts in mouse islets under palmitate treatment with or without NERP-4 ( n = 4 biological replicates). b Snat2 mRNA amounts in 10-week-old db/+ mouse and db/db mouse islets ( n = 4 biological animals). c Snat2 mRNA amounts in islets from db/db mice administered NERP-4 for two weeks ( n = 5 biological replicates). d NERP-4 was administered at 24 and 48 h after the start of siSCR treatment or Snat2 knockdown to MIN6-K8 cells under palmitate ( e – g ) or at 48 h under cytokines ( h ). e OCR ( n = 3 biological replicates). The P values indicated the differences of basal respiration (1.6–14.8 min), ATP production (21.4–34.6 min), and maximal respiration (41.2–54.4 min) in palmitate-treated si Snat2 -MIN6-K8 cells with or without NERP-4. f ATP production ( n = 4 biological replicates). g ROS production ( n = 4 biological replicates). h Cell viability ( n = 6 biological replicates). Representative results of two independent experiments ( a – c , e – h ). Data are mean ± s.e.m. ( a – c , e – h ). One-way ANOVA and Tukey’s multiple comparisons test ( a , e – h ). Unpaired two-tailed Student’s t -test ( b , c ). i Schematic of NERP-4 roles in pancreatic β cells. NERP-4 is processed from VGF, a granin protein that is critical for granule biogenesis in pancreatic β cells. NERP-4 is packed with insulin in secretory granules and secreted by glucose. NERP-4 binds to SNAT2 to stimulate amino acid uptake into β cells, thereby enhancing mitochondrial ATP production, glucose-induced Ca 2+ mobilisation into β cells, and GSIS. NERP-4 expression is reduced in β cells in db/db mice. NERP-4 protects β cells from glucolipotoxicity by reducing ROS production and ER stress, thereby enhancing mitochondrial biogenesis and dynamics. Source data are provided as a file.

Article Snippet: Human SNAT2/SLC38A2 (Myc-DDK–tagged)-CMV 6-entry vector was purchased from Origene (RC201892).

Techniques: Two Tailed Test, Expressing

Journal: eLife

Article Title: PAX3-FOXO1 transgenic zebrafish models identify HES3 as a mediator of rhabdomyosarcoma tumorigenesis

doi: 10.7554/eLife.33800

Figure Lengend Snippet:

Article Snippet: Transfected construct ( Homo sapiens ) , CMV-HES3 (MYC and FLAG tagged) , , , Origene , , Origene:RC224630 , , .

Techniques: Mutagenesis, Injection, Transfection, Construct, Western Blot, Staining, Recombinant, Plasmid Preparation, Generated, Cloning, Sequencing, In Situ, Stripping Membranes, Software