Ontarget sirnas

1C‐TAZ‐WT, 1C‐TAZ‐4SA, 1C‐TAZ‐4SA‐mNLS were kind gifts from Andras Kapus. pBabe‐puro‐HA‐EV, and pBabe‐puro‐HA‐YAP‐5SA were kind gifts from Alex Hergovich. pLenti‐CMV‐SLC7A11‐sh926R‐FLAG‐IRES‐Hygro was purchased from Addgene (# 118702). To generate pSuper‐retro‐puro‐shYAP/TAZ construct, the oligonucleotide 5′‐TGTGGATGAGATGGATACA‐3′ which targets both YAP and TAZ mRNAs was cloned into pSuper‐retro‐puro vector. To generate pGL4.10‐SLC7A11, a DNA fragment from −1,000 bp to +200 bp containing the SLC7A11 promoter was cloned into the pGL4.10 vector.
On‐target siRNAs were purchased from Horizon Discovery and are listed in Appendix Table S1. Antibodies used are listed in Appendix Table S2, and PCR primers are listed in Appendix Table S3.

A cDNA construct encoding for Flag-HIF1α was amplified from a cDNA library and cloned into pcDNA4.0, and Myc-USP29 was amplified from a cDNA library and cloned into pcDNA4/TO/myc-His B. To generate pBabe-USP29, a cDNA fragment coding for USP29 was cloned into pBabe-retro-puro-empty vector. To generate Myc-USP29-CA, Myc-USP29 was mutated at C294S and H831N. pGL4.42 and pRL-CMV were purchased from Promega. On-target siRNAs were purchased from Horizon Discovery. siUSP29#1, siUSP29#2 were ordered from Microsynth and are listed in Suppl. Table II. Myc-USP29-R was mutated on Myc-USP29 to be resistant to siUSP29#1, primers are listed in Suppl. Table II. The Sequences of siRNAs are presented in Suppl. Table II, antibodies used are listed in Suppl. Table III, and oligonucleotides are listed in Suppl. Table IV.

Dermal fibroblasts or endothelial cells from dcSSc patients were treated with 10 µM of LATS kinase inhibitor TRULI/Lats-IN-1 (MedChenExpress HY-138489) or YAP/TEAD inhibitor verteporfin (Cayman Chemical 17334) 0.1–10 µM for 48 to 72 h. Gene knockdown was done using Accell siRNAs (Dharmacon) in dermal fibroblasts and OnTarget siRNAs (Dharmacon) in dermal ECs, following protocols recommended by the manufacturer.