Mass spectrometry data and RNA sequencing data of the effect of Gstp1 on SHR-VSMCs

RNA sequencing analysis was performed to recapitulate possible changes in transcriptome profile caused by downregulation of Gstp1 in SHR-VSMCs. Total RNA was extracted from the cells using Trizol reagent. The concentration, quality and integrity were determined using a NanoDrop spectrophotometer (ThermoFisher Scientific, MA, USA). Total RNA required for single library construction was more than 1 μg. Sequencing libraries were generated using the TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA, USA). Library sequencing was performed on an Illumina Novaseq 6000 platform (Illumina, San Diego, CA, USA).

Mass spectrometry analysis was performed to identify Gstp1 binding proteins in SHR-VSMCs. Proteins were immunoprecipitated using anti-Gstp1 antibody and incubated with Protein A/G PLUS-agarose beads for 4 h. Elution buffer (0.1M glycine–HCl) was used to elute antigen from antibody and 1M Tris–HCl was used for neutralization. After trypsin digestion, peptides were desalted using C18 StageTip(Thermo Fisher Scientific, OH, USA), and vacuum dried. The samples were analyzed by LC-MS/MS using an Easy nLC 1200 coupled to a QExactive HF mass spectrometer (Thermo Scientific, OH, USA). Sequence database searching and protein quantification raw mass spectrometry data were analyzed using the MaxQuant software suite (version 1.6.1.0).