Anti cd317 pe

Human MSCs derived from the umbilical cord and expanded with chemically defined medium [10 (link)] were prepared for cell purification at passage 3. The MSCs were detached with TrypLE and stained with anti-CD317-PE (Thermo Fisher Scientific) or IgG-PE. Then, the CD317⁺ and CD317⁻ MSCs were purified with the BD FACSAria SORP cell sorter (BD Biosciences). Total RNA sequencing was performed at BGI (Beijing Genomics Institute) as described previously [10 (link)]. For shRNA construction, target sequences were cloned and inserted into the lentivirus pLKO.1-puro vector as described previously [32 (link)]. Lentivirus production and cell infection were performed as described previously [8 (link)]. Target sequences are listed in Additional file 1: Table S1.

Cell preparation, antibody staining and flow cytometry were performed as described previously [8 (link)–10 (link), 31 (link)]. Briefly, the MSCs were detached with TrypLE, resuspended in PBS plus 5% BSA (bovine serum albumin, Sigma) and incubated with anti-CD317-PE (Thermo Fisher Scientific), anti-CD73-FITC, anti-CD90-FITC, anti-CD105-FITC, anti-CD45-FITC, anti-CD34-FITC, anti-CD19-FITC, anti-CD11b-FITC, anti-HLADR-FITC, IgG-PE or IgG-FITC (all from BD Biosciences). Data were collected with BD AccuriC6 Plus (BD Biosciences) and analyzed with FlowJo software.