Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the proliferation of T24 cells. (A) RT-qPCR analysis of si-ARAF transfection efficiency. (B and C) Western blot analysis of si-ARAF transfection efficiency. (D) EdU fluorescence assay showing proliferating cells. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Quantitative RT-PCR, Transfection, Western Blot, Fluorescence

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on apoptosis of T24 cells. (A and B) Flow cytometry analysis of apoptosis levels. (C and D) Protein expression of cleaved-caspase 3 and total caspase 3. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. **p<0.01, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Flow Cytometry, Expressing

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the metastatic capacity of T24 cells. (A and B) Quantification of migrating cells. (A and C) Quantification of invading cells. (D and E) Expression of EMT-related proteins in T24 cells. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Expressing

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the p38 MAPK pathway in T24 cells. (A) The level of p-p38, p38, p-ERK, ERK, p-MEK, MEK in T24 cells was determined by western blot assay. (B) p-p38/p38 ratio. (C) p-ERK/ERK ratio. (D) p-MEK/MEK ratio. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Western Blot

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin activated p38 MAPK pathway in si-ARAF transfected T24 cells. (A) The level of p-p38, p38, p-ERK, ERK, p-MEK, MEK in T24 cells was determined by western blot assay. (B) p-p38/p38 ratio. (C) p-ERK/ERK ratio. (D) p-MEK/MEK ratio. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Transfection, Western Blot

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin reversed the effect of ARAF on T24 cell proliferation and apoptosis. (A) EdU fluorescence for cell proliferation capacity. (B) The apoptotic rate of T24 cells. (C and D) The protein level of Cleaved-caspase3 and Caspase3. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Fluorescence

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin reversed the effect of ARAF on T24 cell metastasis and EMT. (A–C) The migratory and invasive ability of T24 cells. (D–F) The protein level of E-cadherin and N-cadherin. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. **p<0.01, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques:

Journal: Advanced Science

Article Title: Piezo1 Channel Mediates Mechanically Programmable Drug Delivery to Potentiate Intravesical Chemotherapy

doi: 10.1002/advs.202522936

Figure Lengend Snippet: Mechanical pressure enhances mechano‐sensitivity of urinary epithelial cells. (A) The scheme of mechanical pressure model in vitro. B) CCK‐8 assay of SV‐HUC, T24, 253J cells exposed on gradient pressure. (C) CCK‐8 assay of cells exposed on 40 cmH 2 O pressure for different times. (D) EdU assay of cells exposed on 40 cmH 2 O pressure for 12 h. (E) F‐Actin staining of SV‐HUC, T24, 253J cells exposed on 40 cm H 2 O pressure for 12 h. (F) The quantitative analysis of EdU assay. (G) The quantitative analysis of nucleo‐cytoplasmic ratio. (H) Expression level of Piezo1/ITGB1/YAP/α‐SMA in cells exposed on gradient pressure. (I) Wound closure assay of SV‐HUC cells exposed on gradient pressure for 24, 48 h. (J) The quantitative analysis of wound closure assay in SV‐HUC cells. (K) The quantitative analysis of wound closure assay in T24 cells. (L) TEM image of SV‐HUC cell exposed on 40 cm H 2 O pressure. (M) The detection process of nanoindentation before and after HP for 2 h. N) Detection of Young's modulus in T24 cells using nanoindentation before and after HP for 2 h. (O) The quantitative analysis of Z position and Young's modulus detection. All data are presented as the Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001. ns, no significant difference.

Article Snippet: The human bladder cancer cell lines T24, 253J, and the normal control cell SV‐HUC, which was the SV‐40 immortalized human uroepithelial cell line, were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: In Vitro, CCK-8 Assay, EdU Assay, Staining, Expressing, Wound Closure Assay

Journal: Advanced Science

Article Title: Piezo1 Channel Mediates Mechanically Programmable Drug Delivery to Potentiate Intravesical Chemotherapy

doi: 10.1002/advs.202522936

Figure Lengend Snippet: Mechanical pressure amplifies calcium influx via Piezo1/ITGB1‐mediated membrane tension. (A) RNA expression level of Piezo1/ITGB1 in cells exposed on mechanical pressure. (B) Protein expression level of Piezo1/ITGB1 in cells exposed on hydrostatic pressure. (C) The heatmap of differentially expressed genes (DEGs) in T24 cells from RNA sequencing. (D) Go analysis of DEGs in T24 cells from RNA sequencing. E) KEGG analysis of DEGs in T24 cells from RNA sequencing. (F) The flow cytometry result of intracellular calcium flux in T24 and SV‐HUC cells exposed on hydrostatic pressure for 0, 0.5, 1, 2, and 4 h using Fluo‐4AM probe. (G) The flow cytometry result of intracellular calcium flux in T24 cells exposed on hydrostatic pressure with Piezo1 agonists or inhibitors. (H) The image of intracellular calcium flux in T24 cells exposed on hydrostatic pressure with agonists or inhibitors using Fluo‐4AM probe. (I) Expression level of Piezo1/ITGB1 signal of cells exposed on pressure with agonists or inhibitors. (J) Membrane tension through fluorescence lifetime imaging (FLIM) in T24 cells exposed on pressure for 2 h. All data are presented as the Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001. ns, no significant difference.

Article Snippet: The human bladder cancer cell lines T24, 253J, and the normal control cell SV‐HUC, which was the SV‐40 immortalized human uroepithelial cell line, were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Membrane, RNA Expression, Expressing, RNA Sequencing, Flow Cytometry, Fluorescence, Imaging

Journal: Advanced Science

Article Title: Piezo1 Channel Mediates Mechanically Programmable Drug Delivery to Potentiate Intravesical Chemotherapy

doi: 10.1002/advs.202522936

Figure Lengend Snippet: Mechanical pressure enhances DOX accumulation in tumor. (A) Proliferation assay of SV‐HUC, T24, 253J cells exposed on 40 cm H 2 O pressure and different concentrations of DOX. (B) The uptake efficiency of DOX in cells exposed on HP from flow cytometry. (C) The quantitative analysis of the apoptosis rate of cells induced by DOX and HP from flow cytometry. D)The quantitative result of DOX uptake efficiency from flow cytometry using a series of endocytic inhibitors. (E) DOX uptake efficiency of T24 cells exposed on DOX from flow cytometry using a series of endocytic inhibitors. (F) DOX uptake efficiency of T24 cells exposed on DOX and HP from flow cytometry using a series of endocytic inhibitors. (G) DOX accumulation in tumor spheres exposed on HP and different concentrations of DOX. H) Immunofluorescence of Piezo1/ITGB1 in tumor spheres exposed on HP and DOX. I) The quantitative analysis of DOX intensity in tumor spheres treated with different concentrations of DOX. J) The quantitative analysis of Piezo1 and ITGB1 expression levels in tumor spheres. K) The quantitative analysis of DOX accumulation in tumor spheres treated with DOX and HP. All data are presented as the Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001. ns, no significant difference.

Article Snippet: The human bladder cancer cell lines T24, 253J, and the normal control cell SV‐HUC, which was the SV‐40 immortalized human uroepithelial cell line, were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Proliferation Assay, Flow Cytometry, Immunofluorescence, Expressing

Journal: Advanced Science

Article Title: Piezo1 Channel Mediates Mechanically Programmable Drug Delivery to Potentiate Intravesical Chemotherapy

doi: 10.1002/advs.202522936

Figure Lengend Snippet: Mechanical pressure enhances tumor apoptosis induced by chemical drugs. (A) Western blotting results of Piezo1/ITGB1 expression level in five matched pairs of bladder normal and tumor tissues. (B) CCK‐8 assay of T24 cell exposed on 40 cm H 2 O pressure and different concentrations of MMC. (C) Immunofluorescence of Piezo1/ITGB1 in SV‐HUC cells exposed on HP and DOX. (D) The quantitative analysis of Piezo1/ITGB1 expression and DOX nuclear accumulation in SV‐HUC cells. (E) Immunofluorescence of Piezo1/ITGB1 in T24 cells exposed on HP and DOX. (F) The quantitative analysis of Piezo1/ITGB1 expression and DOX nuclear accumulation in T24 cells. (G,H) Expression level of mechanical axis and apoptosis proteins in cells treated with DOX and mechanical pressure. (I) Expression level of apoptosis proteins in T24 cells treated with MMC and mechanical pressure.

Article Snippet: The human bladder cancer cell lines T24, 253J, and the normal control cell SV‐HUC, which was the SV‐40 immortalized human uroepithelial cell line, were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Western Blot, Expressing, CCK-8 Assay, Immunofluorescence

Journal: Advanced Science

Article Title: Piezo1 Channel Mediates Mechanically Programmable Drug Delivery to Potentiate Intravesical Chemotherapy

doi: 10.1002/advs.202522936

Figure Lengend Snippet: Piezo1 mediates mechanical potentiation of chemotherapeutic efficacy. (A) Proliferation assay of overexpressed cells exposed on HP and different concentrations of DOX. (B) Expression level of the mechanical pathway and apoptosis proteins in cells treated with DOX and HP. (C) The apoptosis rate induced by DOX and HP from flow cytometry. (D) The quantitative analysis of apoptosis rate from flow cytometry. (E) The flow cytometry result of DOX accumulation in T24 cells exposed on HP with Piezo1 agonists or inhibitors. (F) Expression level of the mechanical pathway and apoptosis proteins in Sh‐Piezo1 cells treated with DOX and HP. (G) The flow cytometry result of DOX accumulation in Sh‐Piezo1 cells exposed on HP. (H) The flow cytometry result of DOX uptake in T24 cells treated with YAP‐specific inhibitor verteporfin. All data are presented as the Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001. ns, no significant difference.

Article Snippet: The human bladder cancer cell lines T24, 253J, and the normal control cell SV‐HUC, which was the SV‐40 immortalized human uroepithelial cell line, were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Proliferation Assay, Expressing, Flow Cytometry