Journal: Reproductive biomedicine online
Article Title: Transcriptomic analysis supports collective endometrial cell migration in the pathogenesis of adenomyosis
doi: 10.1016/j.rbmo.2022.05.007
Figure Lengend Snippet: Validation of potential target genes in top enriched biological processes in the eutopic endometrium of women with versus without adenomyosis. (A) mRNA levels of IL1B , IL18 and TNF in eutopic endometrium of adenomyosis patients ( n = 10) and controls ( n = 10) detected by quantitative reverse transcription polymerase chain reaction analysis (qRT-PCR). All endometrial samples were in the proliferative phase. β -ACTIN was used as reference gene for qRT-PCR analysis. P -values are 0.002, 0.041 and 0.022, respectively; (B) mRNA levels of MMP1 , MMP8 , MMP13 and TIMP1 in eutopic endometrium of adenomyosis patients ( n = 10) and controls ( n = 10) detected by qRT-PCR analysis. P -values are 0.002, 0.001, 0.008 and 0.0085, respectively; (C) mRNA levels of COL1A1 , COL1A2 and COL3A1 in eutopic endometrium of adenomyosis patients (n = 10) and controls (n = 10) detected by qRT-PCR analysis; (D) mRNA levels of HAS1 , HAS2 , HAS3 and CD44 in eutopic endometrium of adenomyosis patients ( n = 10) and controls ( n = 10) detected by qRT-PCR analysis. P -values are 0.016, 0.049, 0.015 and 0.484, respectively. Data are presented as means ± SEM. * P < 0.05, ** P < 0.01; (E) protein levels of COL1A1, COLIII, MMP1 and MMP13 in the endometrium of adenomyosis patients and controls. n = 6 for each group. GAPDH was used as reference control, and the ratio of band intensity of a target protein to that of the intensity of GAPDH was obtained at each target protein level . The P -value of the protein level of COL1A1, COLIII, MMP1 and MMP13 in adenomyosis versus controls are 0.009, 0.024, 0.018 and 0.031, respectively. * P < 0.05, ** P < 0.01; (F) Masson stain of eutopic endometrium in adenomyosis patients versus controls.
Article Snippet: Samples of protein (35 μg) were separated on 10% sodium dodecyl sulfate gels in 80–120 V for 1.5 h and then wet transferred to a nitrocellulose membrane at 200 mA for 1.5–2 h. After blocking in 5% bovine serum albumin for 1 h, the nitrocellulose membrane was incubated with primary antibody dilution buffer (Beyotime, Shanghai, China) diluted antibody against matrix metalloproteinase 1 (MMP1) (1:1000) (Proteintech, Wuhan, China), MMP13 (1:1000) (Proteintech, Wuhan, China), collagen III (COLIII) (1:1000) (Proteintech, Wuhan, China), and COL1A1 (1:1000) (Cell Signaling Technology, Danvers, MA, USA) at 4°C overnight.
Techniques: Biomarker Discovery, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Control, Staining
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