human rcc cell lines a498  (ATCC)

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a EHMT2 expression in normal and RCC samples derived from TCGA database. P values were calculated using Student’s t -test (** P < 0.01). b Kaplan‒Meier plot showing that the overall survival rates of patients with low EHMT2 expression were substantially higher than those of patients with high EHMT2 expression in RCC tissues. P values were calculated using Student’s t -test (*** P < 0.001). c Immunohistochemical staining for EHMT2. Kidney cancer tissues were purchased from TissueArray ( https://www.tissuearray.com ). Scale bar, 200 μm. d DAVID-based GO analysis of the RNA-seq results from the siEHMT2 (#1) and siCont groups, which included 1207 DEGs. e , f Cell growth assay after transfection with siEHMT2 and siCont for 48 h. e A498 and Caki-1 cells were fixed with 100% methanol and stained with the CV solution. Scale bar, 500 μm. f CCK-8 solution was added to the culture medium and the cells were incubated for 5 min at 37 °C. Cell growth was measured using a microplate reader (450 nm). The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001). g Western blot analysis of cells transfected with siEHMT2 transfection using anti-EHMT2, anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 and Caki-1 cells. h FACS analysis of Annexin V staining was performed after the cells were transfected with siEHMT2 or siCont. The lower right and upper right quadrants indicate early apoptotic cells and late apoptotic cells, respectively (top). Quantification of apoptosis: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (bottom). i FACS analysis using the Muse Caspase-3/7 working solution was performed after the cells were transfected with siEHMT2 or siCont. The upper right image shows the proportions of apoptotic and dead cells (top). Quantification of caspase-3/7 activity: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (bottom).

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Expressing, Derivative Assay, Immunohistochemical staining, Staining, RNA Sequencing, Growth Assay, Transfection, CCK-8 Assay, Incubation, Western Blot, Control, Activity Assay

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a Migration and invasion assays were performed using the A498 and Caki-1 cell lines after EHMT2 knockdown. Cell migration and invasion assays were performed after 24 h (A498) and 48 h (Caki-1). The migrating/invading cells were stained with CV. Scale bar, 500 μm (left). Quantification of migrating/invading cells: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (right). b Migration assay of A498 and Caki-1 cells after treatment with TGF-β. The cell migration assay was performed after 24 h (A498) and 48 h (Caki-1). The migrating cells were stained with CV. Scale bar, 500 μm (left). Quantification of migrating cells: the data are presented as the means ± SDs of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001) (right). c RT‒qPCR analysis of E-cadherin and N-cadherin expression in cells transfected with siEHMT2. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, ** P < 0.01, *** P < 0.001). d Migration and invasion assays were performed using the A498 and Caki-1 cell lines after treatment with TGF-β and EHMT2 knockdown. Cell migration and invasion assays were performed after 24 h (A498) and 48 h (Caki-1). The migrating/invading cells were stained with CV. Scale bar, 500 μm (left). Quantification of migrating/invading cells: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t- tests (** P < 0.01, *** P < 0.001) (right). e RT‒qPCR analysis of E-cadherin and N-cadherin expression in cells transfected with siEHMT2. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001).

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Migration, Knockdown, Staining, Cell Migration Assay, Expressing, Transfection

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a , b Cell growth assay after treatment with BIX for 48 h: A498 and Caki-1 cells were fixed with 100% methanol and stained with a CV solution, scale bar, 500 μm ( a ); CCK-8 solution was added to the culture medium and the cells were incubated for 5 min at 37 °C. Cell growth was measured using a microplate reader (450 nm) ( b ). The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001). c Western blot analysis of cells treated with BIX using anti-EHMT2, anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 and Caki-1 cells. d FACS analysis of Annexin V staining was performed after BIX treatment. The lower right and upper right quadrants indicate early apoptotic cells and late apoptotic cells, respectively (left). Quantification of apoptosis: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (right). e FACS analysis using the Muse Caspase-3/7 working solution was performed after BIX treatment. The upper right image shows the proportions of apoptotic and dead cells (left). Quantification of caspase-3/7 activity: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (right). f Migration and invasion assays were performed in A498 and Caki-1 cells after BIX treatment. Cell migration and invasion assays were performed after 24 h (A498) and 48 h (Caki-1). The migrating/invading cells were stained with CV. Scale bar, 500 μm (left). Quantification of migrating/invading cells: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001) (right). g Migration and invasion assays were performed after the A498 and Caki-1 cell lines were treated with TGF-β and BIX. Cell migration and invasion assays were performed after 24 h (A498) and 48 h (Caki-1). The migrating/invading cells were stained with CV. Scale bar, 500 μm (left). Quantification of migrating/invading cells. The data are presented as the means ± s.d. of three independent experiments: P values were calculated using Student’s t -tests (* P < 0.05, *** P < 0.001) (right). h RT‒qPCR analysis of E-cadherin and N-cadherin expression in cells after BIX treatment. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, *** P < 0.001).

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Growth Assay, Staining, CCK-8 Assay, Incubation, Western Blot, Control, Activity Assay, Migration, Expressing

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a A heat map of RNA-seq data from siEHMT2- and siCont-transfected cells. b RNA-seq results for DDIT3 expression after EHMT2 knockdown. c RT‒qPCR analysis of DDIT3 expression in cells transfected with siEHMT2. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001). d Correlation analysis of the expression of the EHMT2 and DDIT3 genes derived from TCGA portal and using analysis of variance (ANOVA). e Immunohistochemical staining for EHMT2 and DDIT3. Kidney cancer tissues were purchased from TissueArray ( https://www.tissuearray.com ). Scale bar, 200 μm. f Immunocytochemical staining for DDIT3. A498 and Caki-1 cells transfected with siEHMT2 and siCont were fixed with 100% methanol and stained with an anti-DDIT3 antibody (Alexa Fluor 488, green) and DAPI (blue). Scale bar, 150 μm. g Graphical abstract of the ChIP primer design for the DDIT3 promoter region. h The ChIP assay was performed with an anti-H3K9me2 antibody. The result is shown as relative enrichment compared to the control in A498 and Caki-1 cells after siEHMT2 transfection. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, ** P < 0.01).

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: RNA Sequencing, Transfection, Expressing, Knockdown, Derivative Assay, Immunohistochemical staining, Staining, Control

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a Immunocytochemical staining for DDIT3. A498 and Caki-1 cells were treated with BIX, fixed with 100% methanol and stained with an anti-DDIT3 antibody (Alexa Fluor 488, green) and DAPI (blue). Scale bar, 300 μm. b The ChIP assay was performed with an anti-H3K9me2 antibody. The results are shown as relative enrichment compared to the control in A498 and Caki-1 cells after BIX treatment. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, ** P < 0.01, *** P < 0.001). c Cell growth assay after cotransfection with siDDIT3 and siEHMT2 for 48 h. A498 and Caki-1 cells were fixed with 100% methanol and stained with a CV solution. Scale bar, 500 μm (top). CCK-8 solution was added to the culture medium and the cells were incubated for 5 min at 37 °C. Cell growth was measured using a microplate reader (450 nm). The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (bottom). d FACS analysis of Annexin V staining was performed after cells were cotransfected with siDDIT3 and siEHMT2. Quantification of apoptosis: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001). e FACS analysis using the Muse Caspase-3/7 working solution was performed after cells were cotransfected with siDDIT3 and siEHMT2. Quantification of caspase-3/7 activity: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001). f Western blot analysis of cells cotransfected with siDDIT3 and siEHMT2 using anti-EHMT2, anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 and Caki-1 cells.

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Staining, Control, Growth Assay, Cotransfection, CCK-8 Assay, Incubation, Activity Assay, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a Cell growth assay after treatment with Fb7-311 for 24 h. A498 and Caki-1 cells were fixed with 100% methanol and stained with a CV solution. Scale bar, 500 μm (top). CCK-8 solution was added to the culture medium and the cells were incubated for 5 min at 37 °C. Cell growth was measured using a microplate reader (450 nm). The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (bottom). b FACS analysis of Annexin V staining was performed after cells were treated with Fb7-311. The lower right and upper right quadrants indicate early apoptotic cells and late apoptotic cells, respectively (top). Quantification of apoptosis: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (bottom). c FACS analysis using the Muse Caspase-3/7 working solution was performed after cells were treated with Fb7-311. The upper right image shows the proportions of apoptotic and dead cells (top). Quantification of caspase-3/7 activity: the data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, *** P < 0.001) (bottom). d RT‒qPCR analysis of EHMT2 and DDIT3 expression after cells were treated with Fb7-311. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, ** P < 0.01, *** P < 0.001). e Western blot analysis of cells treated with Fb7-311 using anti-EHMT2, anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 and Caki-1 cells. f Immunocytochemical staining for EHMT2 and DDIT3. A498 and Caki-1 cells were treated with Fb7-311 fixed with 100% methanol and stained with an anti-DDIT3 antibody (Alexa Fluor 488, green) and DAPI (blue). Scale bar, 150 μm. g The ChIP assay was performed with an anti-H3K9me2 antibody on the DDIT3 promoter region. The result is shown as relative enrichment compared to the control in A498 and Caki-1 cells after Fb7-311 treatment. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (** P < 0.01). h Cell growth assay after treatment with indole-3-carbinol for 72 h. A498 cells were fixed with 100% methanol and stained with a CV solution. Scale bar, 500 μm (left). CCK-8 solution was added to the culture medium and the cells were incubated for 5 min at 37 °C. Cell growth was measured using a microplate reader (450 nm). The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001) (right). i RT‒qPCR analysis of EHMT2 and DDIT3 expression after cells were treated with indole-3-carbinol. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (** P < 0.01).

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Growth Assay, Staining, CCK-8 Assay, Incubation, Activity Assay, Expressing, Western Blot, Control

Journal: Experimental & Molecular Medicine

Article Title: Gut microbiota modulation of epigenetic target EHMT2: Lacticaseibacillus rhamnosus Fb7-311 regulated renal cell carcinoma apoptosis and metastasis

doi: 10.1038/s12276-026-01659-6

Figure Lengend Snippet: a The 3D spheroid formation assay. The cells transfected with siEHMT2 and siCont were loaded onto ULA plates and incubated for 48 h. The cells were photographed under a microscope each day. Scale bar, 500 μm. b Western blot analysis of cells after EHMT2 knockdown using anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 cell. c RT‒qPCR analysis of EHMT2 and DDIT3 expression after EHMT2 knockdown. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (* P < 0.05, ** P < 0.01, *** P < 0.001). d The 3D spheroid formation assay. Cells cotransfected with siEHMT2 and siDDIT3 were loaded onto ULA plates and incubated for 48 h. The cells were photographed under a microscope each day. Scale bar, 500 μm. e Western blot analysis of cells cotransfected with siEHMT2 and siDDIT3 using anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 cell. f RT‒qPCR analysis of EHMT2 and DDIT3 expression in cells cotransfected with siEHMT2 and siDDIT3. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001). g The 3D spheroid formation assay. After BIX was added to ULA plates, the cells were incubated for 48 h. The cells were then photographed under a microscope each day. Scale bar, 500 μm. h Western blot analysis of cells treated with BIX using anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 cell. i RT‒qPCR analysis of DDIT3 expression in cells treated with BIX. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -test (*** P < 0.001). j The 3D spheroid formation assay. The cells were loaded onto ULA plates after treatment with Fb7-311 and incubated for 48 h. The cells were photographed under a microscope each day. Scale bar, 500 μm. k Western blot analysis of Fb7-311-treated cells using anti-PARP and anti-ACTB antibodies. ACTB was used as the internal control in A498 cell. l RT‒qPCR analysis of EHMT2 and DDIT3 expression in Fb7-311-treated cells. The data are presented as the means ± s.d. of three independent experiments. P values were calculated using Student’s t -tests (** P < 0.01, *** P < 0.001). m , n BIX treatment suppressed the growth of xenograft tumors in nude mice. Both the control and BIX were intraperitoneally injected three times a week after A498 cell implantation: tumor volumes ( P values were calculated using two-way ANOVA (** P < 0.01)) (m) and macroscopic image of tumors on day 24 ( n ). o Representative H&E-stained mouse tumor sections. Scale bars, 200 μm. p Immunohistochemical staining for DDIT3 in mouse tumor sections. Scale bar, 200 μm.

Article Snippet: The human RCC cell lines A498 and Caki-1 were purchased from the Korean Cell Line Bank and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C.

Techniques: Tube Formation Assay, Transfection, Incubation, Microscopy, Western Blot, Knockdown, Control, Expressing, Injection, Staining, Immunohistochemical staining

Journal: Biochemistry and Biophysics Reports

Article Title: Sorafenib induces ferroptosis in human renal cell carcinoma cells through CCAT/enhancer-binding protein homologous protein

doi: 10.1016/j.bbrep.2025.102143

Figure Lengend Snippet: Sorafenib induces ferroptosis in RCC. (A) Cell viability was analyzed by CCK8 assay in A498 cells treated with DMSO (control) or various concentrations of sorafenib for 24, 48, and 72 h (n = 3/group). (B) Cell viability analyzed by CCK8 assay in A498 cells treated with DMSO (control), 20 μM sorafenib, and sorafenib in combination with a ferroptosis inhibitor (Ferrostatin-1: Fer-1) or activator (RSL3) for 24 h (n = 3/group). (C) Representative histograms of the fluorescence intensity of C11-oxidized membrane lipids in A498 cells treated with DMSO (control), 20 μM sorafenib, and sorafenib with Fer-1 for 24 h and a graph summarizing the mean fluorescence intensity of those cells (n = 3/group). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 between the indicated groups. Data are presented as the mean ± SD.

Article Snippet: A human A498 RCC cell line was purchased from the Korean Cell Line Bank.

Techniques: CCK-8 Assay, Control, Fluorescence, Membrane

Journal: Biochemistry and Biophysics Reports

Article Title: Sorafenib induces ferroptosis in human renal cell carcinoma cells through CCAT/enhancer-binding protein homologous protein

doi: 10.1016/j.bbrep.2025.102143

Figure Lengend Snippet: Sorafenib induces ER stress in RCC. (A) Cell viability was analyzed by CCK8 assay of A498 cells treated with DMSO (control), 20 μM sorafenib, and sorafenib in combination with an ER stress inhibitor for 24 h (n = 3/group). (B) Representative histograms of the fluorescence intensity of C11-oxidized membrane lipids in A498 cells treated with DMSO (control), 20 μM sorafenib, and sorafenib with an ER stress inhibitor for 24 h and a graph summarizing the mean fluorescence intensity of those cells (n = 3/group). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 between the indicated groups. Data are presented as the mean ± SD.

Article Snippet: A human A498 RCC cell line was purchased from the Korean Cell Line Bank.

Techniques: CCK-8 Assay, Control, Fluorescence, Membrane

Journal: Biochemistry and Biophysics Reports

Article Title: Sorafenib induces ferroptosis in human renal cell carcinoma cells through CCAT/enhancer-binding protein homologous protein

doi: 10.1016/j.bbrep.2025.102143

Figure Lengend Snippet: Sorafenib upregulates ATF4 and CHOP in RCC. (A) Relative GRP78 mRNA levels were quantified by RT-qPCR in A498 cells treated with DMSO or 20 μM sorafenib for 24, 48 and 72 h (n = 3/group). (B) Relative mRNA levels of ATF4, ATF6, and XBP1s were quantified by RT-qPCR in A498 cells treated with DMSO or 20 μM sorafenib for 24 h (n = 3/group). (C and D) Relative mRNA levels of ATF4 (C) and CHOP (D) were quantified by RT-qPCR in A498 cells treated with DMSO or 20 μM sorafenib at different time points (n = 3/group). (E) Grp78, ATF6, XBP1, ATF4 and CHOP protein levels were analyzed by Western blot in A498 cells treated with DMSO or 20 μM sorafenib for 24 h (n = 3/group). The intensity of bands was quantified using ImageJ. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 between the indicated group. Data are presented as the mean ± SD.

Article Snippet: A human A498 RCC cell line was purchased from the Korean Cell Line Bank.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Sorafenib induces ferroptosis in human renal cell carcinoma cells through CCAT/enhancer-binding protein homologous protein

doi: 10.1016/j.bbrep.2025.102143

Figure Lengend Snippet: CHOP contributes to sorafenib-induced ferroptosis through inhibition of SLC7A11 in RCC. (A) Relative CHOP mRNA levels were quantified by RT-qPCR in CHOP siRNA knockdown A498 cells, followed by treatment with DMSO or sorafenib (SORA) 24 h after siRNA transfection (n = 3/group). (B) CHOP protein levels were analyzed by Western blot in A498 cells treated with DMSO or sorafenib with or without CHOP knockdown (n = 3/group). (C) Cell viability was analyzed by CCK8 assay in A498 cells treated with DMSO or sorafenib with or without CHOP knockdown (n = 3/group). (D) Representative histograms of the fluorescence intensity of C11-oxidized membrane lipids in A498 cells treated with DMSO or sorafenib with or without CHOP knockdown and a graph summarizing the mean fluorescence intensity of those cells (n = 3/group). (E) Relative mRNA levels of SLC7A11 were quantified by RT-qPCR in A498 cells treated with DMSO or sorafenib with or without CHOP knockdown (n = 3/group). (F) SLC7A11 protein levels were analyzed by Western blot in A498 cells treated with DMSO or sorafenib with or without CHOP knockdown (n = 3/group). (G) Kaplan-Meier survival curves of overall survival according to SLC7A11 expression in the TCGA renal clear cell carcinoma cohorts. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 between the indicated groups. Data are presented as the mean ± SD.

Article Snippet: A human A498 RCC cell line was purchased from the Korean Cell Line Bank.

Techniques: Inhibition, Quantitative RT-PCR, Knockdown, Transfection, Western Blot, CCK-8 Assay, Fluorescence, Membrane, Expressing

Journal: Frontiers in Oncology

Article Title: Overexpression of BNIP3 in renal carcinoma cells can promote apoptosis of renal carcinoma cells through HIF-1α-BNIP3-mediated autophagy

doi: 10.3389/fonc.2025.1614378

Figure Lengend Snippet: Expression levels of BNIP3 and HIF-1α in renal cell carcinoma (RCC) cell lines. (A) Western blot analysis of BNIP3 and HIF-1α expression levels in A498,786-O, CAKI-1, ACHN, and GRC cell lines. (B, C) Western blot analysis of BNIP3 expression levels in A498 and 786-O cell lines. (D) mRNA expression levels of BNIP3 in A498 and 786-O cell lines were examined using reverse transcription-quantitative PCR. Data are mean ± SD, * P < 0.05, ** P < 0.01.

Article Snippet: Human RCC cell lines A498, 786-O, CAKI-1, ACHN, and GRC were purchased from KeyGEN Company (Nanjing, China).

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Frontiers in Oncology

Article Title: Overexpression of BNIP3 in renal carcinoma cells can promote apoptosis of renal carcinoma cells through HIF-1α-BNIP3-mediated autophagy

doi: 10.3389/fonc.2025.1614378

Figure Lengend Snippet: Overexpression of BNIP3 promotes the dissociation of the Bcl-2/Beclin1 complex through competitive binding of Bcl-2 under hypoxia conditions. (A, B) The dissociation of Bcl-2/Beclin1 complex in BNIP3-overexpressing A498 and 786-O cells was indicated by the Co-IP experiment with Beclin1 antibody and Bcl-2 antibody.

Article Snippet: Human RCC cell lines A498, 786-O, CAKI-1, ACHN, and GRC were purchased from KeyGEN Company (Nanjing, China).

Techniques: Over Expression, Binding Assay, Co-Immunoprecipitation Assay

Journal: Frontiers in Oncology

Article Title: Overexpression of BNIP3 in renal carcinoma cells can promote apoptosis of renal carcinoma cells through HIF-1α-BNIP3-mediated autophagy

doi: 10.3389/fonc.2025.1614378

Figure Lengend Snippet: Overexpression of BNIP3 promotes autophagy in renal cancer cells under hypoxia. (A) Immunofluorescence staining was used to detect the expression levels of LC3B (×40). (B) Mean fluorescence intensity of LC3B. (C) The autophagosomes in A498 and 786-O cells were visualized using transmission electron microscopy (TEM) at a magnification of ×20000 after exposure to hypoxia. (D-F) Western blot detection of LC3-I, LC3-II, and p62. (G, H) Overexpression of BNIP3 affecting A498 and 786-O cells ROS. (I) Concentration of mitochondrial membrane permeability transition pore (mPTP). Data are mean ± SD, * P < 0.05, ** P < 0.01.

Article Snippet: Human RCC cell lines A498, 786-O, CAKI-1, ACHN, and GRC were purchased from KeyGEN Company (Nanjing, China).

Techniques: Over Expression, Immunofluorescence, Staining, Expressing, Fluorescence, Transmission Assay, Electron Microscopy, Western Blot, Concentration Assay, Membrane, Permeability

Journal: Frontiers in Oncology

Article Title: Overexpression of BNIP3 in renal carcinoma cells can promote apoptosis of renal carcinoma cells through HIF-1α-BNIP3-mediated autophagy

doi: 10.3389/fonc.2025.1614378

Figure Lengend Snippet: Overexpression of BNIP3 promotes apoptosis of cancer cells by mediating autophagy under hypoxia. (A, B) Apoptosis was detected by flow cytometry. (C, D) Overexpression of BNIP3 affecting A498 and 786-O cells ROS. (E-J) Western blot detection of LC3-I, LC3-II, p62, caspase3, cleaved caspase3 and Bax. Data are mean ± SD, * P < 0.05, ** P < 0.01, # P < 0.05, ## P < 0.01.

Article Snippet: Human RCC cell lines A498, 786-O, CAKI-1, ACHN, and GRC were purchased from KeyGEN Company (Nanjing, China).

Techniques: Over Expression, Flow Cytometry, Western Blot