Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus, female) | Hsd: Athymic Nude-Foxn1nu | Envigo | Autosomal recessive mutation on nu locus on chromosome 11; T-cell deficient; accepts xenograft transplantation. | |
| Cell line (Homo sapiens) | HUVEC | This paper | Umbilical cord endothelial cells isolated in house; see materials and methods section | |
| Cell line (Homo sapiens) | HSaVEC | PromoCell | # C-12231 | Saphenous vein endothelial cells |
| Cell line (Homo sapiens) | HPF-c | PromoCell | # C-12360 | Pulmonary fibroblast |
| Transfected construct (Homo sapiens) | LV-PIK3CA | This paper | PIK3CAwt | Lentiviral construct to transfect and express PIK3CA wild-type; produced using Addgene plasmid #116771 |
| Transfected construct (Homo sapiens) | LV-PIK3CA-H1047R | This paper | PIK3CAH1047R | Lentiviral construct to transfect and express PIK3CA with oncogenic point mutation on p.H1047R; produced using Addgene plasmid #116500 |
| Biological sample (Homo sapiens) | Patients’ AST/VM tissue | This paper | Patients’ lesion | Tissue material collected during surgery; see materials and methods section |
| Biological sample (Homo sapiens) | Endothelial cells from patients’ AST/VM | This paper | Patient EC (CD31+) | Freshly isolated from AST/VM tissue; see materials and methods section |
| Biological sample (Homo sapiens) | Stromal cells from patients’ AST/VM | This paper | Patient SC (CD31-/ Vimentin+) | Freshly isolated from AST/VM tissue; see materials and methods section |
| Recombinant DNA reagent | pHAGE-PIK3CA | Addgene | # 116771 | Plasmid encoding PIK3CA wild-type |
| Recombinant DNA reagent | pHAGE-PIK3CA-H1047R | Addgene | # 116500 | Plasmid encoding PIK3CA with oncogenic point mutation on p.H1047R |
| Sequence-based reagent | siRNA: nontargeting control #1 | Thermo Fisher Scientific | # 4390844 | Silencer Select siRNA |
| Sequence-based reagent | siRNA: nontargeting control #2 | Thermo Fisher Scientific | # 4390847 | Silencer Select siRNA |
| Sequence-based reagent | siRNA: targeting to human TGFA | Thermo Fisher Scientific | Assay ID: s14053 | Silencer select siRNA |
| Sequence-based reagent | PrimePCR ddPCR mutation detection assays for PIK3CA c.3140A>G (p.H1047R) | Bio-Rad | Assay IDs: dHsaMDM5225715851 (mut) and dHsaMDW5225715853 (wt) | |
| Sequence-based reagent | PrimePCR ddPCR mutation detection assays for PIK3CA c.3140A>T (p.H1047L) | Bio-Rad | Assay IDs: dHsaMDM2916088171 (mut) and dHsaMDW2916088173 (wt) | |
| sequence-based reagent | PrimePCR ddPCR mutation detection assays for PIK3CA c.1633G>A (p.E545K) | Bio-Rad | Assay IDs: dHsaMDM9869636521 (mut) and dHsaMDW9869636523 (wt) | |
| sequence-based reagent | PrimePCR ddPCR mutation detection assays for PIK3CA c.1624G>A (p.E542K) | Bio-Rad | Assay IDs: dHsaMDM3010833491 (mut) and dHsaMDW3010833493 (wt) | |
| sequence-based reagent | Custom-design Taqman SNP Genotyping assays for TEK c.2740C>T (p.L914F) | Thermo Fisher Scientific | # 4331349 | fwd 5’-CTTCCCTCCAGGCTACTT-3’, rev 5’-AATGCTGGGTCCGTCT-3’, reporter 1 (HEX) 5’-CTTGCGAAGGAAGTCCAGAAGGTTTC-3’, and reporter 2 (FAM) 5’- CTTGCGAAAGAAGTCCAGAAGGTTTC-3’ |
| peptide, recombinant protein | rhTGFA | Sigma-Aldrich | # GF313 | |
| commercial assay or kit | Human CD31 microbead kit | Miltenyi Biotec | # 130-091-935 | |
| commercial assay or kit | RNeasy Mini Kit | Qiagen | # 74106 | |
| commercial assay or kit | Human Quantikine TGFA ELISA | R&D Systems | # DTGA00 | |
| commercial assay or kit | Human Quantikine VEGF ELISA | R&D Systems | # DVE00 | |
| chemical compound, drug | Afatinib | MedChem Express | # HY-10261 | |
| software, algorithm | SproutAngio |
https://github.com/mbbio/SproutAngio (Beter et al., 2023) |
Open access tool for quantitation |