Fig 1: Validation of hub genes in the darkmagenta module. (A and B) ROC curves of NFASC (A) and CHL1 (B) for distinguishing ectopic endometrial samples from non-ectopic endometrial samples. (C and D) Relative protein expressions of NFASC (C) and CHL1 (D) in the Ctrl, Eu and Ec, as determined by IHC staining. Scale Bars = 50 µm. **p < 0.01, ***p < 0.001.
Fig 2: The relationship between hub genes and infiltrating immune cells. For each hub gene, samples were divided into two groups (high expression or low expression group) based on median gene expression. (A and B) The stacked bar chart shows the infiltration ratio of 22 types of immune cells in each sample of in the high- or low-NFASC (A) and high- or low-CHL1 (B) groups, respectively. (C) Box plot shows the infiltration difference of immune cells between the high- and low-NFASC groups. (D) Box plot shows the infiltration difference of immune cells between the high- and low-CHL1 groups. (E and F) The relationship between NFASC (E) or CHL1 (F) and immune infiltrating cells, respectively.
Fig 3: Effects of CHL1 silencing on immortalized but non-neoplastic mammary cellsA. HBL-100 cells were transduced with pHIV1-SIREN+scramble, pHIV1-SIREN+shCHL1_1, or pHIV1-SIREN+shCHL1_2 and selected with puromycin for 11 days. CHL1 silencing efficiency was checked by western blot, using a-tubulin as a loading control. Numbers indicate the amount of CHL1 relative to that of a-tubulin, as measured by densitometry. B. Cell proliferation was measured by RTCA for 5 additional days upon CHL1 silencing. C. Effects of CHL1 knockdown on cell migration were measured for 24 h. Images were acquired at 50x magnification with NIS-Elements, using an Olympus BX51 microscope (Olympus, Tokyo, Japan). D. Cell invasion was measured by RTCA for 3 additional days after CHL1 silencing.
Fig 4: Epigenetic status of CHL1 in BC patientsThe methylation of three CpG sites in the CHL1 gene promoter was interrogated by pyrosequencing in a series of 142 BC cases, 45 adjacent-to-tumour tissues, and 19 non-neoplastic mammary tissues from reduction mammoplasties. The horizontal lines in each group represent the median of the series. (***, p < 0.001).
Fig 5: CHL1 protein expression in BCImmunohistochemistry was employed to measure CHL1 expression in 57 paired breast tumour and adjacent-to-tumour samples, along with 20 non-neoplastic tissues from reduction mammoplasties. It was scored as: 0, no expression; 1: weak expression; 2: intermediate expression; and 3: strong expression (***, p < 0.001; *, p < 0.05). Images were acquired at 400x magnification using a Leica DMD 108 digital microscope (Leica, Wetzlar, Germany).
Supplier Page from Abcam for Anti-CHL1 antibody