Journal: Bioactive Materials

Article Title: Mesenchymal stromal cells-loaded 3D radially aligned composite scaffold with potentiated paracrine signaling for sequential bone regeneration

doi: 10.1016/j.bioactmat.2026.02.059

Figure Lengend Snippet: Temporal analysis of the BMSC paracrine profile on different scaffolds. (A) Confocal microscopy images from Live/Dead fluorescence staining of BMSCs encapsulated within the PCL/HAp-GelMA/BMSCs scaffold after 1, 3, 5, and 14 d of 3D culture (live cells, green; dead cells, red). (B) The concentrations of key paracrine factors (TGF-β, PGE2, VEGF, HGF, and BMP-2) from BMSCs cultured in different scaffolds, quantified from culture supernatants at day 3 and day 7. (C) Corresponding relative mRNA expression levels of TGFB1, PTGS2, VEGFA, HGF, and BMP-2 in BMSCs at day 3 and day 7, as determined by qPCR analysis. Data are presented as mean ± SD (n = 3) *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns: not significant.

Article Snippet: ELISA kits for PGE2 (Cat. No. E-EL-0034), TGF-β (Cat. No. E-EL-0162), VEGF (Cat. No. E-EL-R2603), and HGF (Cat. No. E-EL-R0496) were purchased from Elabscience (Wuhan, China).

Techniques: Confocal Microscopy, Fluorescence, Staining, Cell Culture, Expressing

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: TGF-β induces EMT differentiation and upregulates lncRNA XIST expression in RC cells. 786-O and Caki-1 cells were treated with 10 ng/mL TGF-β for 0, 7, 14, and 21 days. (A) Cell morphology was assessed by light microscopy at each time point (magnification, 400×). (B) XIST gene expression was assessed by RT-qPCR in the 786-O and Caki-1 cells. *, P<0.05. EMT, epithelial-mesenchymal transition; lncRNA, long non-coding RNA; RC, renal cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor beta; XIST, X inactive-specific transcript.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Expressing, Light Microscopy, Gene Expression, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: LncRNA XIST silencing inhibits proliferation, EMT, migration, and invasion, and induces apoptosis in TGF-β-mediated RC cells. (A) 786-O and Caki-1 cells were treated with 10 ng/mL TGF-β for 21 days and then transfected with si-XIST-1, si-XIST-2, or si-XIST-3. The silencing effect of XIST in the TGF-β-treated RC cells was analyzed by RT-qPCR. (B) The morphology of the TGF-β-treated RC cells was observed after interfering with XIST by microscopy (magnification, 400×). (C) XIST expression was quantified by RT-qPCR in the cells treated with TGF-β, si-XIST, or their combination. (D) Cell proliferation was assessed by MTT assay. (E) Apoptosis was evaluated by flow cytometry. (F) N-cadherin, Vimentin, and Snail expression was assessed by western blot analysis. PVDF membranes were pre-cut prior to antibody incubation to enable the simultaneous detection of these proteins from the same lysates. Complete, uncut membrane images are provided in the supplementary file (available at https://cdn.amegroups.cn/static/public/tau-2026-0536-1.pdf ). GAPDH served as a loading control on a separate membrane strip. (G) E-cadherin expression was assessed by IF staining (magnification, 200×). (H,I) Migration (H) and invasion (I) was assessed by Transwell assays in the TGF-β-treated RC cells following XIST silencing (crystal violet staining; magnification, 200×). *, P<0.05; **, P<0.01. NC: PBS + si-NC; si-XIST: PBS + si-XIST; TGF-β: TGF-β + si-NC. EMT, epithelial-mesenchymal transition; FITC, fluorescein isothiocyanate; IF, immunofluorescence; lncRNA, long non-coding RNA; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium; bromideNC, negative control; OD570, optical density at 570 nm; PBS, phosphate-buffered saline; PI, propidium iodide; PVDF, polyvinylidene difluoride; RC, renal cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si, small-interfering RNA; TGF-β, transforming growth factor beta; XIST, X inactive-specific transcript.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Migration, Transfection, Quantitative RT-PCR, Microscopy, Expressing, MTT Assay, Flow Cytometry, Western Blot, Incubation, Membrane, Control, Stripping Membranes, Staining, Immunofluorescence, Negative Control, Saline, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: LncRNA XIST targets and regulates miR-141-3p. (A) Predicted binding sites between XIST and miR-141-3p were identified by bioinformatic analysis. (B) XIST and miR-141-3p expression levels were assessed by RT-qPCR following XIST knockdown in TGF-β-treated 786-O and Caki-1 cells. (C) The luciferase activity of WT- or Mut-XIST in miR-141-3p-overexpressing 293T cells was assessed by dual-luciferase reporter assays. *, P<0.05. LncRNA, long non-coding RNA; Mut, mutant; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor beta; WT, wild type; XIST, X inactive-specific transcript.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Knockdown, Luciferase, Activity Assay, Mutagenesis, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: Inhibition of miR-141-3p attenuates the suppressive effects of lncRNA XIST silencing on proliferation, EMT, migration, and invasion, while promoting apoptosis in TGF-β-treated RC cells. TGF-β-treated 786-O and Caki-1 cells were transfected with si-XIST and/or a miR-141-3p inhibitor. (A) XIST and miR-141-3p expression was assessed by RT-qPCR assay. (B) The morphology of RC cells was observed by light microscopy (magnification, 400×). (C) Cell proliferation was assessed by MTT assay. (D) Cell apoptosis was assessed by flow cytometry. (E) N-cadherin, Vimentin, and Snail protein expression levels were examined by western blot analysis. PVDF membranes were pre-cut prior to antibody incubation to enable the simultaneous detection of these proteins from the same lysates. Complete, uncut membrane images are provided in the supplementary file (available at https://cdn.amegroups.cn/static/public/tau-2026-0536-1.pdf ). GAPDH served as a loading control on a separate membrane strip. (F) E-cadherin expression was assessed by IF staining (magnification, 200×). (G,H) Cell migration and invasion were assessed by Transwell assays (crystal violet staining; magnification, 200×). *, P<0.05; **, P<0.01; ***, P<0.001. NC: inhibitor NC + si-XIST. EMT, epithelial-mesenchymal transition; FITC, fluorescein isothiocyanate; IF, immunofluorescence; lncRNA, long non-coding RNA; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium; NC, negative control; PI, propidium iodide; PVDF, polyvinylidene difluoride; RC, renal cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si, small-interfering RNA; TGF-β, transforming growth factor beta; XIST, X inactive-specific transcript.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Inhibition, Migration, Transfection, Expressing, Quantitative RT-PCR, Light Microscopy, MTT Assay, Flow Cytometry, Western Blot, Incubation, Membrane, Control, Stripping Membranes, Staining, Immunofluorescence, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: miR-141-3p targets and regulates ZEB1. (A) Potential binding sites between miR-141-3p and ZEB1 were predicted using TargetScanHuman 7.1. (B,C) After miR-141-3p overexpression, miR-141-3p and ZEB1 expression levels were analyzed by RT-qPCR (B) and western blot analysis (C) in TGF-β-treated 786-O and Caki-1 cells. PVDF membranes were pre-cut prior to antibody incubation to enable the simultaneous detection of these proteins from the same lysates. Complete, uncut membrane images are provided in the supplementary file (available at https://cdn.amegroups.cn/static/public/tau-2026-0536-1.pdf ). GAPDH served as a loading control on a separate membrane strip. (D) Changes in the luciferase activity of WT or Mut ZEB1 in the miR-141-3p-overexpressing 293T cells were measured by dual-luciferase reporter gene assays. **, P<0.01. Mut, mutant; NC, negative control; PVDF, polyvinylidene difluoride; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor beta; WT, wild type; ZEB1, zinc-finger E-box binding protein 1.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Binding Assay, Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Incubation, Membrane, Control, Stripping Membranes, Luciferase, Activity Assay, Mutagenesis, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Translational Andrology and Urology

Article Title: Long non-coding RNA (lncRNA) XIST drives TGF-β-induced renal cancer progression via miR-141-3p/ZEB1 signaling

doi: 10.21037/tau-2026-0536

Figure Lengend Snippet: ZEB1 overexpression attenuates the inhibitory effects of miR-141-3p mimics on proliferation, EMT, migration, and invasion, while decreasing apoptosis in TGF-β-treated RC cells. TGF-β-treated 786-O and Caki-1 cells were transfected with miR-141-3p mimic, mimic NC, and/or ZEB1 overexpression plasmids. (A) ZEB1 expression was examined by RT-qPCR. (B) The morphology of RC cells was observed by light microscopy (magnification, 400×). (C) Cell proliferation was assessed by MTT assay. (D) Cell apoptosis was assessed by flow cytometry. (E) ZEB1, N-cadherin, Vimentin, and Snail expression levels were detected by western blot analysis. PVDF membranes were pre-cut prior to antibody incubation to enable the simultaneous detection of these proteins from the same lysates. Complete, uncut membrane images are provided in the supplementary file (available at https://cdn.amegroups.cn/static/public/tau-2026-0536-1.pdf ). GAPDH served as a loading control on a separate membrane strip. (F) E-cadherin expression was assessed by IF staining (magnification, 200×). (G,H) Cell migration and invasion were evaluated by Transwell assays (crystal violet staining; magnification, 200×). NC: mimics NC; 141 mimics: miR-141-3p mimics. *, P<0.05. EMT, epithelial-mesenchymal transition; IF, immunofluorescence; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium; NC, negative control; OD570, optical density at 570 nm; PVDF, polyvinylidene difluoride; RC, renal cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor beta; ZEB1, zinc-finger E-box binding protein 1.

Article Snippet: For TGF-β stimulation, the 786-O and Caki-1 cells were treated with 10 ng/mL recombinant TGF-β (catalog #RP00161; ABCLONAL) for 0, 7, 14, and 21 days.

Techniques: Over Expression, Migration, Transfection, Expressing, Quantitative RT-PCR, Light Microscopy, MTT Assay, Flow Cytometry, Western Blot, Incubation, Membrane, Control, Stripping Membranes, Staining, Immunofluorescence, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Binding Assay