Angiopoietin-like Protein 7/ANGPTL7 Protein 4960-AN-025 from R&D Systems, a Bio-Techne Brand

Published Figures

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Communications biology
October 03 2022

Fig 1: Expression analysis of ANGPTL7 Gln175His, Arg177*, and Trp188* in a HEK293 cell line.a, b Western blotting shows intra- and extra-cellular protein levels of ANGPTL7 wild-type (WT), Gln175His, Arg177*, and Trp188*. c ELISA was run to quantify intra- and extra-cellular protein levels of ANGPTL7 WT, Gln175His, Arg177*, and Trp188* transiently transfected in HEK293 cells (whole-cell lysate was diluted 1:1,000; supernatant was diluted 1:10,000. Both whole-cell lysate and supernatant were normalized against the total amount of protein from the whole-cell lysate); ****= P < 1 × 10–4 statistical difference from WT whole cell lysate; ^ = P < 0.05, ^^^ = P < 1 × 10–4 statistical difference from WT supernatant. d Ratio of secreted versus intracellular ANGPTL7 WT, Gln175His, Arg177*, and Trp188* protein levels. Raw ANGPTL7 WT, Gln175His, Arg177*, and Trp188* protein levels were normalized to the whole-lysate protein concentration; **** = P < 1 × 10-4 statistical difference from WT. Western blotting and ELISA analysis were repeated on three independent biological replicates. Technical replicates (n = 3) were run for the ELISA analysis. P values were calculated by one-way ANOVA with Tukey’s post hoc analysis. All data are presented as mean and error bars indicate the standard error of the mean (SEM). MWt molecular weight marker.
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Communications biology
October 03 2022

Fig 2: ANGPTL7 expression in ocular tissues across species.RNA-sequencing-based expression levels (measured in transcripts per million, TPM, and represented as median and interquartile range) are highest in cornea, trabecular meshwork (TM), and sclera in human (a), and African green monkey (b) eyes, and in cornea, TM, sclera, optic nerve head, and choroid/RPE in C57BL/6 J mouse eyes (c); In situ hybridization (RNAscope) shows ANGPTL7/Angptl7 (red) expression in TM, cornea and sclera in human (d) and murine (e) eyes. Scale bars represent 100 µm. DAPI staining (blue) counterstains cell nuclei. RPE retinal pigmented epithelium, CB ciliary body, SC Schlemm’s canal, CM ciliary muscle, AC anterior chamber, RGC retinal ganglion cell, INL inner nuclear layer, ONL outer nuclear layer.
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Communications biology
October 03 2022

Fig 3: Association of ANGPTL7 variants with glaucoma.a Meta-analysis results for Gln175His with glaucoma across 8 different cohorts. b Cross-ancestry meta-analysis of Arg177* and Trp188* across 5 European (EUR) and 3 African (AFR) ancestry cohorts. The variants in the meta-analysis of EUR and AFR cohorts were Arg177* and Trp188*, respectively. GHS Geisinger DiscovEHR, UKB UK Biobank, SINAI Mt. Sinai Medical School BioMe Biobank, MALMO Malmö Diet and Cancer Study, EstBB the Estonia Biobank at the University of Tartu, HUNT the HUNT study from Nord-Trøndelag, CGPS-CCHS the Copenhagen General Population Study and the Copenhagen City Heart Study, POAAGG Primary Open Angle African-American Glaucoma Genetics, MAF Minor allele frequency.
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Communications biology
October 03 2022

Fig 4: In situ characterization of Angptl7 mRNA in WT and Angptl7 KO mouse eyes.Angptl7 mRNA was not expressed in any ocular tissue in Angptl7 KO mice whereas it was expressed in TM, cornea, and sclera of WT mice as shown by in situ hybridization (RNAscope). Brightfield images showing following probes. a Negative control: DapB. b Positive control: Ubc (red). c WT mice: Angptl7 (red), and (d) Angptl7 KO mice: Angptl7 (no signal). Scale bars represent 100 µm.
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Communications biology
October 03 2022

Fig 5: Increasing mAngptl7 levels in mouse eyes increases IOP.Murine Angptl7 (mAngptl7) protein was injected into mouse eyes via intravitreal (a) or intracameral route (b), and IOP was measured over time. After an initial drop, IOP remained elevated for several days in mAngptl7-treated eyes compared to PBS treated (CTRL (PBS)) eyes. * = P < 0.05; ** = P < 0.01 statistical significance compared to PBS treatment. Statistical analyses were performed using Student’s t test.