1) FCS files from spectral flow cytometry experiments performed on AML patient blasts. 2) HDR donor for pSTAT1 to add the degron tag and mCherry expression.

From methods section of the article:AML patient blast samples were obtained from the Vanderbilt University Medical Center Hematology Tissue Repository. One portion containing 400,000 cells was split into two conditions: 1) PBS control and 2) IFNγ-stimulated for 15 minutes. After stimulation, cells were fixed in 1.6% PFA and permeabilized in ice-cold methanol. Barcoding was performed using Pacific Orange (PO) (ThermoFisher, P30253) at two distinct concentrations (high: 100 ng/mL, low: 12 ng/mL). The barcoded cells were washed, pooled, and stained with an antibody cocktail (table S7). Spectral flow cytometry was conducted using the Cytek Aurora (Cytek Biosciences) to measure key surface marker expression and STAT1 phosphorylation.A cloning plasmid (IDT, pUCIDT-AMP) was modified to serve as the homology-directed repair (HDR) donor by incorporating the STAT1 sequence from the cut site to the stop codon, an FKBP12F36V degron tag, an HA-tag, a P2A sequence, and an mCherry fluorescent protein sequence. These elements were flanked by a 5′ homology arm (200 bp) and a 3′ homology arm (450 bp). The HDR donor plasmid map is available on Figshare.本文方法部分内容如下：急性髓系白血病（Acute Myeloid Leukemia, AML）患者原始细胞样本取自范德堡大学医学中心血液科组织库。取一份含400,000个细胞的样本分为两个处理组：1）磷酸盐缓冲液（Phosphate Buffered Saline, PBS）对照组；2）干扰素γ（Interferon γ, IFNγ）刺激15分钟组。刺激完成后，将细胞用1.6%多聚甲醛（Paraformaldehyde, PFA）固定，随后置于冰浴甲醇中进行透化处理。采用太平洋橙染料（Pacific Orange, PO）、赛默飞世尔科技（ThermoFisher）货号P30253，以两种不同浓度（高浓度：100 ng/mL，低浓度：12 ng/mL）完成条形码标记。标记后的细胞经洗涤、混合后，使用抗体鸡尾酒（详见补充表S7）进行染色。采用赛泰克生物科学（Cytek Biosciences）的Cytek Aurora流式细胞仪开展光谱流式细胞术检测，以检测关键表面标志物的表达水平及信号转导与转录激活因子1（Signal Transducer and Activator of Transcription 1, STAT1）的磷酸化状态。 本研究对克隆质粒（整合DNA技术, IDT，pUCIDT-AMP）进行改造，将其作为同源定向修复（Homology-Directed Repair, HDR）供体载体：插入从酶切位点至终止密码子的STAT1序列、FKBP12F36V降解标签、血凝素（Hemagglutinin, HA）标签、P2A序列以及mCherry荧光蛋白序列。上述功能元件两侧分别带有5'同源臂（200 bp）与3'同源臂（450 bp）。该同源定向修复供体质粒的图谱可于Figshare平台获取。