Sybr green premix

A total of 27 EC tissues and 20 normal endometrial tissues were obtained from patients who underwent a hysterectomy or endometrial curettage for endometrial-irrelevant diseases in Shengjing Hospital of China Medical University, China, from 2017 to 2019. All patients provided informed consent, and this study was approved by the Ethics Committee of Shengjing Hospital of China Medical University. Histological diagnosis and tumor grade were assessed by three experienced pathologists in accordance with the International Federation of Gynecology and Obstetrics (FIGO 2009). None of the patients received any local or systemic treatment preoperatively.
Total RNA of EC tissues and normal endometrial tissues were extracted with TRIzol reagent (Vazyme, Nanjing, China). The synthesis of cDNAs corresponding to the mRNAs of interest depended on PrimeScript RT-polymerase (Vazyme). SYBR-Green Premix (Vazyme) with specific PCR primers (Sangon Biotech Co., Ltd., Shanghai, China). Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. The 2-ΔΔCt method was used to calculate fold-changes. Primer sequences are shown in Supplementary Table 1. The difference of the expression of E2Fs in tumor tissues and normal tissues were evaluated with Student’s t-test in GraphPad Prism7 software (GraphPad, Inc., La Jolla, CA, USA).

We obtained 52 breast cancer tissues and normal breast tissues from patients who underwent a breast cancer removal in Union Hospital of Tongji Medical College. The studies involving human participants were reviewed and approved by the Ethics Committee of Union Hospital of Tongji Medical College. The patients/participants provided their written informed consent to participate in this study. Histological diagnosis and tumor grade were assessed by three experienced pathologists in accordance with the 2010 American Joint Committee on Cancer staging system.
The total RNA of breast cancer tissues and normal tissues was extracted with the TRIzol reagent (Vazyme, Nanjing, China). The synthesis of cDNAs corresponding to the mRNAs of interest depended on PrimeScript RT-polymerase (Vazyme) and SYBR-Green Premix (Vazyme) with specific PCR primers (Sangon Biotech Co., Ltd, Shanghai, China). The specific primers used were as follows: GAPDH-F: GGG​AAG​GTG​AAG​GTC​GGA​GT; GAPDH-R: GGG​GTC​ATT​GAT​GGC​AAC​A; BCL2-F: ACT​GGC​TCT​GTC​TGA​GTA​AG; BCL2-R: CCT​GAT​GCT​CTG​GGT​AAC. The fold-changes were calculated with the 2^−ΔΔCt method. The difference in BCL2 expression and the prognosis of BCL2 in breast cancer were evaluated with Student’s t-test and the Kaplan–Meier analysis in GraphPad Prism 7 software (GraphPad, Inc., La Jolla, CA, United States).

Total RNA was extracted using TRIzol reagent (Vazyme, Nanjing, China). Subsequently, cDNAs were synthesized using Prime Script RT-polymerase (Vazyme). SYBR Green Premix (Vazyme) with specific PCR primers (Sangon Biotech, Shanghai, China) were used to detect the expression level of corresponding gene RNA. The primer sequences are provided in Supplementary Table 2. The fold-changes were calculated using the 2^−ΔΔCT method.

Total RNA was extracted from tissues using TRIzol reagent (Vazyme, Nanjing, China). PrimeScript RT-polymerase (Vazyme) was used to reverse-transcribe cDNAs corresponding to the mRNAs of interest. qRT-PCR was performed using SYBR-Green Premix (Vazyme) with specific PCR primers (Sangon Biotech Co., Ltd, Shanghai, China). Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. The 2^−ΔΔCt method was used to calculate fold-changes. Primer sequences are listed in Additional file 1: Table S1.

Total RNA was isolated from cell samples utilizing RNA-easy isolation reagent (Vazyme Biotech), which was then reversed into cDNA by PrimeScript Mix reagent (Takara). SYBR Green Premix (Vazyme Biotech) was prepared for use in the PCR system. The value of individual genes was finally standardized to the GAPDH expression level. The primer sequences for indicated genes are displayed in Supplementary Table S1.

Total cell RNA was extracted by RNA isolation reagent (Takara), then reversed into cDNA by PrimeScript Mix reagent (Takara). SYBR Green Premix (Vazyme biotech) was utilized for PCR reaction system. The value of individual genes was standardized to GAPDH expression level. Supplementary Table S2 displays primer sequences of all genes.