Total RNA of 1-3 million sarcoma cells was isolated using the High Pure RNA Isolation Kit (Roche, Basel, Switzerland) and cDNA synthesis was done using FastGeneScriptase II (NIPPON Genetics Europe, Düren, Germany) according to the manufacturer’s instructions. Primer sequences for MICA, MICB, ULBP1-4 and GAPDH were used as previously described (24 (link), 25 (link)) (Table S4). Reverse transcriptase–polymerase chain reaction (RT-PCR) was performed as described previously (26 (link)). Quantitative PCR (q-PCR) was performed using PerfeCTa SYBR Green FastMix (Quanta Biosciences Beverly, MA) with a LightCycler 480 (Roche) instrument.
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