Journal: Apoptosis

Article Title: Single-cell profiling and machine learning identify cuproptosis-related fibroblast subpopulations and fibrogenesis modulator AEBP1 in endometriosis

doi: 10.1007/s10495-026-02342-x

Figure Lengend Snippet: FDX1 knockdown attenuates copper ionophore-associated mitochondrial alterations and profibrotic responses in endometrial stromal cells. ThESCs were transfected with negative-control siRNA (siNC) or FDX1 siRNA (siFDX1), followed by treatment with CuCl 2 plus elesclomol (Cu + ele) or vehicle, as indicated. A Representative western blots and densitometric quantification of FDX1, LIAS, and Lip-DLAT. Tubulin was used as the loading control. B Representative JC-1 flow cytometry plots and quantification of JC-1 green fluorescence. C Representative MitoSOX staining images and quantification of mitochondrial ROS levels. Nuclei were counterstained with DAPI. D Representative immunofluorescence staining of AEBP1 and FDX1, with quantification of relative fluorescence intensity. E Representative western blots and densitometric quantification of AEBP1, α-SMA, and CTGF. F Representative western blots and densitometric quantification of β-catenin and c-Myc. Western blot quantification was derived from three independent experiments. MitoSOX fluorescence was quantified from three independent microscopic fields per group. Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple-comparisons test. ** P < 0.01; **** P < 0.0001. Scale bars = 50 μm. si NC, negative-control small interfering RNA; si FDX1, FDX1-specific small interfering RNA; Cu + ele, CuCl 2 plus elesclomol; α-SMA, alpha-smooth muscle actin; CTGF, connective tissue growth factor; ROS, reactive oxygen species

Article Snippet: Human endometrial stromal cells (ThESCs) were obtained from the American Type Culture Collection (ATCC; catalog no. CRL-4003) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) enriched with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA).

Techniques: Knockdown, Transfection, Negative Control, Western Blot, Control, Flow Cytometry, Fluorescence, Staining, Immunofluorescence, Derivative Assay, Small Interfering RNA

Journal: Apoptosis

Article Title: Single-cell profiling and machine learning identify cuproptosis-related fibroblast subpopulations and fibrogenesis modulator AEBP1 in endometriosis

doi: 10.1007/s10495-026-02342-x

Figure Lengend Snippet: AEBP1 modulates profibrotic marker expression and β-catenin pathway-related proteins in endometrial stromal cells. ThESCs were transfected with AEBP1-specific siRNA (siAEBP1) or AEBP1 overexpression plasmid (ovAEBP1), followed by treatment with CuCl 2 plus elesclomol (Cu + ele) or vehicle, as indicated. A Representative immunofluorescence staining of AEBP1 and α-SMA in siNC, siAEBP1, Cu + ele, and siAEBP1 + Cu + ele groups, with quantification of relative fluorescence intensity. B Representative western blots and densitometric quantification of AEBP1, α-SMA, and CTGF in siNC, siAEBP1, Cu + ele, and siAEBP1 + Cu + ele groups. C Representative western blots and densitometric quantification of AEBP1, α-SMA, and CTGF in ovNC, Cu + ele, and ovAEBP1 groups. D Representative immunofluorescence staining of AEBP1 and β-catenin in siNC, siAEBP1, Cu + ele, and siAEBP1 + Cu + ele groups, with quantification of relative fluorescence intensity. E Representative western blots and densitometric quantification of β-catenin and c-Myc in siNC, siAEBP1, Cu + ele, and siAEBP1 + Cu + ele groups. F Representative western blots and densitometric quantification of β-catenin and c-Myc in ovNC, Cu + ele, and ovAEBP1 groups. Tubulin was used as the loading control for all western blot analyses. Western blot quantification was derived from three independent experiments. Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple-comparisons test. ns, not significant; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bars = 50 μm. si NC, negative-control small interfering RNA; si AEBP1, AEBP1-specific small interfering RNA; ov NC, empty vector control; ovAEBP1, AEBP1 overexpression plasmid; α-SMA, alpha-smooth muscle actin; CTGF, connective tissue growth factor

Article Snippet: Human endometrial stromal cells (ThESCs) were obtained from the American Type Culture Collection (ATCC; catalog no. CRL-4003) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) enriched with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA).

Techniques: Marker, Expressing, Transfection, Over Expression, Plasmid Preparation, Immunofluorescence, Staining, Fluorescence, Western Blot, Control, Derivative Assay, Negative Control, Small Interfering RNA