In the original publication of Juraleviciute et al., (2020), the description of two methods utilized in a study “Quantitative real-time PCR” and “siRNA knockdown” was missing from the Supporting Information Data S1 file at the time of submission. These methods should read as below.

Quantitative real-time PCR: Total RNA was extracted from cell samples and tissues using a Macherey-Nagel NucleoSPin RNA extraction kit (Duren, Germany) according to the manufacturer's instruction. Reverse transcription reactions were performed with SuperScript VILO cDNA Synthesis Kit (cat.no 11754050; Invitrogen) using random primers and following the manufacturer's protocol. Total amount of 0.5 µg RNA used in 20 µl of reaction mixture. Real-time PCR analyses were performed using TaqMan Fast Universal PCR Master Mix (2×) and TaqMan Gene Expression Assays (MX2—Hs01550811_m1; MX1—Hs00895608_m1; GUSB—HS99999908_m1; Applied Biosystems). 25 µl of PCR mixture contained 0.5 µl cDNA, 250 nM TaqMan probe, and 900 nM of each primer. RT-qPCRs were performed on a QuantStudio™ 5 Real-Time PCR system (Applied Biosystems; Thermo Fisher Scientific) running the following program: (1) enzyme activation at 95°C for 20 s and (2) 40 cycles of PCR at 95°C for 1 s and 60°C for 20 s. Relative transcript expression levels were normalized against a housekeeping gene beta-glucuronidase GUSB and calculated using a comparative Ct method. Each sample was run in triplicate. Experiments in cell lines were performed with three biological replicates; results are presented as mean ± SE.

siRNA knockdown: All cells were plated into 6-well plates at density of 2 × 10⁵ per well 24 hr before transfection. Cells were transfected with 20 nM two independent siRNAs targeting MX2. #1 targeting sequence 5′-GGAAACAGGAGCCAACCAAtt-3′ (cat. no. AM16706, ID 11695; Ambion, Life Technologies) and #2—targeting sequence 5′-AUCCACCUGAAUGCCUACUtt-3′ (cat.no. 1299001, ID HSS106817; Invitrogen; Life Technologies). Non-targeting siRNA (cat no. 4390843; Ambion, Life Technologies) was used as a negative control. Transfection performed using Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA) transfection reagent and following manufacturer's protocol. Cells were harvested for protein, RNA, and flow analyses after 48 hr and for cell viability assays after 72 hr.

在 Juraleviciute 等人 ( 2020 )的原始出版物中，提交时的支持信息数据 S1 文件中缺少对“定量实时 PCR”和“siRNA 敲除”研究中使用的两种方法的描述。这些方法应如下所示。

定量实时 PCR：根据制造商的说明，使用 Macherey-Nagel NucleoSPin RNA 提取试剂盒（Duren，德国）从细胞样品和组织中提取总 RNA。使用 SuperScript VILO cDNA Synthesis Kit (cat.no 11754050; Invitrogen) 使用随机引物并遵循制造商的方案进行逆转录反应。20 µl 反应混合物中使用的 0.5 µg RNA 总量。使用 TaqMan Fast Universal PCR Master Mix (2x) 和 TaqMan Gene Expression Assays（MX2—Hs01550811_m1；MX1—Hs00895608_m1；GUSB—HS99999908_m1；Applied Biosystems）进行实时 PCR 分析。25 µl PCR 混合物包含 0.5 µl cDNA、250 nM TaqMan 探针和 900 nM 的每种引物。RT-qPCR 在 QuantStudio™ 5 实时 PCR 系统（Applied Biosystems；Thermo Fisher Scientific）上进行，运行以下程序：(1) 95°C 20 s 酶活化；(2) 95°C 1 s 和60°C 20 s 40 个循环的PCR。相对转录表达水平针对管家基因 β-葡萄糖醛酸酶 GUSB 进行标准化，并使用比较 Ct 方法计算。每个样品一式三份运行。细胞系实验进行了三个生物学重复；结果表示为平均值±SE。

siRNA 敲低：所有细胞在转染前 24 小时以每孔2 × 10 ^{5 的}密度接种到 6 孔板中。细胞用 20 nM 两种靶向MX2 的独立 siRNA 转染。#1 靶向序列 5'-GGAAACAGGAGCCAACCAAtt-3'（目录号 AM16706，ID 11695；Ambion，Life Technologies）和 #2-靶向序列 5'-AUCCACCUGAAUGCCUACUtt-3'（目录号 1299001，ID HSS106817；Invitogen） ；生命科技）。非靶向 siRNA（目录号 4390843；Ambion，Life Technologies）用作阴性对照。使用 Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA) 转染试剂并遵循制造商的方案进行转染。48 小时后收集细胞用于蛋白质、RNA 和流量分析，72 小时后用于细胞活力测定。