a498 cell lines  (ATCC)


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    ATCC a498 cell lines
    A498 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 htb 44 cell lines  (ATCC)


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    ATCC a498 htb 44 cell lines
    A498 Htb 44 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 cell lines  (ATCC)


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    ATCC a498 cell lines
    A498 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 human kidney carcinoma cell line  (ATCC)


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    ATCC a498 human kidney carcinoma cell line
    Induction of cell death by EA in <t>A498</t> RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .
    A498 Human Kidney Carcinoma Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy"

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-32-57

    Induction of cell death by EA in A498 RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .
    Figure Legend Snippet: Induction of cell death by EA in A498 RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .

    Techniques Used: Incubation, Prestoblue Assay, Staining, Flow Cytometry

    Caspases are not activated in-EA induced cell death. A498 cells cells were treated with 100 nM EA or 0.1% DMSO (control) for 43 h, or with 200 μM etoposide for 24 h. Cells were then harvested and stained with the FLICA reagent which only binds active caspases. Levels of active caspase were then determined by fluorescence (A) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-caspase-3 antibody. B-actin was probed as a control for protein loading (B) .
    Figure Legend Snippet: Caspases are not activated in-EA induced cell death. A498 cells cells were treated with 100 nM EA or 0.1% DMSO (control) for 43 h, or with 200 μM etoposide for 24 h. Cells were then harvested and stained with the FLICA reagent which only binds active caspases. Levels of active caspase were then determined by fluorescence (A) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-caspase-3 antibody. B-actin was probed as a control for protein loading (B) .

    Techniques Used: Staining, Fluorescence, Western Blot

    EA induces autophagy in A498 cells. A498 cells were treated with 200 nM EA or 0.1% DMSO (control) for 46 h and with 500 nM rapamycin for 20 h. Autophagy was measured by staining autolysosomes and earlier autophagic compartments with the fluorescent probe Cyto-ID ® Green. Samples were then analyzed in the green (FL1) channel of the FACS Caliber flow cytometer (A) . Cells were treated with 200 nM EA or 0.1% DMSO (control) for 45 h and then stained with Hoechst nuclear stain and Cyto-ID ® Green detection reagent followed by fixing with 4% formaldehyde. The stained cells were then analyzed by fluorescence microscopy. Panels a and c show cells treated with 0.1% DMSO and panels b and d show cells treated with EA. Nuclei are stained in blue. Autolysosomes and earlier autophagic compartments are stained in green (B) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-LC3B antibody. B-actin was probed as a control for protein loading (C) .
    Figure Legend Snippet: EA induces autophagy in A498 cells. A498 cells were treated with 200 nM EA or 0.1% DMSO (control) for 46 h and with 500 nM rapamycin for 20 h. Autophagy was measured by staining autolysosomes and earlier autophagic compartments with the fluorescent probe Cyto-ID ® Green. Samples were then analyzed in the green (FL1) channel of the FACS Caliber flow cytometer (A) . Cells were treated with 200 nM EA or 0.1% DMSO (control) for 45 h and then stained with Hoechst nuclear stain and Cyto-ID ® Green detection reagent followed by fixing with 4% formaldehyde. The stained cells were then analyzed by fluorescence microscopy. Panels a and c show cells treated with 0.1% DMSO and panels b and d show cells treated with EA. Nuclei are stained in blue. Autolysosomes and earlier autophagic compartments are stained in green (B) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-LC3B antibody. B-actin was probed as a control for protein loading (C) .

    Techniques Used: Staining, Flow Cytometry, Fluorescence, Microscopy, Western Blot

    Inhibition of autophagy does not affect EA-induced cell death. A498 cells were cultured in the presence and absence of 100 nM EA or, with EA and 1X NEAA in combination for 44 h. Autophagy was then determined by flow cytometry after staining with Cyto-ID ® (A) . A498 cells were treated with 150 nM EA, 0.1% DMSO, 1X NEAA, 200 μM VP16 or with 100 nM EA plus 1X NEAA for 46 h. Cell viability was then determined using the PrestoBlue ® assay (B) . A498 cells were treated as in (B) and then apoptosis was determined by measuring histone-associated DNA fragments by ELISA (C) .
    Figure Legend Snippet: Inhibition of autophagy does not affect EA-induced cell death. A498 cells were cultured in the presence and absence of 100 nM EA or, with EA and 1X NEAA in combination for 44 h. Autophagy was then determined by flow cytometry after staining with Cyto-ID ® (A) . A498 cells were treated with 150 nM EA, 0.1% DMSO, 1X NEAA, 200 μM VP16 or with 100 nM EA plus 1X NEAA for 46 h. Cell viability was then determined using the PrestoBlue ® assay (B) . A498 cells were treated as in (B) and then apoptosis was determined by measuring histone-associated DNA fragments by ELISA (C) .

    Techniques Used: Inhibition, Cell Culture, Flow Cytometry, Staining, Prestoblue Assay, Enzyme-linked Immunosorbent Assay

    EA blocks the G 2 /M transition of the cell cycle. A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 45 h. The cells were then fixed and stained with PI. The PI content of cells was measured by flow cytometry as described under Methods.
    Figure Legend Snippet: EA blocks the G 2 /M transition of the cell cycle. A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 45 h. The cells were then fixed and stained with PI. The PI content of cells was measured by flow cytometry as described under Methods.

    Techniques Used: Staining, Flow Cytometry

    EA inhibits activation of AKT and ERK kinsases. A498 cells were cultured with 100 nM EA or with 0.1% DMSO (control) for the indicated times and protein was Isolated. Western blot analysis was performed as described under Methods using antibodies against AKT, ERK, and AMPK and their phosphorylated counterparts. B-actin was probed to control for protein loading. (+) control; Jurkat cell extract.
    Figure Legend Snippet: EA inhibits activation of AKT and ERK kinsases. A498 cells were cultured with 100 nM EA or with 0.1% DMSO (control) for the indicated times and protein was Isolated. Western blot analysis was performed as described under Methods using antibodies against AKT, ERK, and AMPK and their phosphorylated counterparts. B-actin was probed to control for protein loading. (+) control; Jurkat cell extract.

    Techniques Used: Activation Assay, Cell Culture, Isolation, Western Blot

    human kidney carcinoma cell line a498  (ATCC)


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    ATCC human kidney carcinoma cell line a498
    Human Kidney Carcinoma Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    kidney epithelial cell line a498  (ATCC)


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    ATCC kidney epithelial cell line a498
    Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line <t>A498.</t> The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.
    Kidney Epithelial Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Vitamin D Induction of the Human Antimicrobial Peptide Cathelicidin in the Urinary Bladder"

    Article Title: Vitamin D Induction of the Human Antimicrobial Peptide Cathelicidin in the Urinary Bladder

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0015580

    Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line A498. The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.
    Figure Legend Snippet: Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line A498. The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Immunohistochemistry, Staining

    cell lines a498  (ATCC)


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    ATCC cell lines a498
    GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in <t>A498</t> and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively
    Cell Lines A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "GABPA-activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma"

    Article Title: GABPA-activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-022-02382-6

    GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in A498 and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively
    Figure Legend Snippet: GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in A498 and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Techniques Used: Expressing, Western Blot, Over Expression, Flow Cytometry

    L-2-HG accumulation induces epigenetic silence of GABPA expression. A GABPA methylation is significantly higher in ccRCC tumors than in corresponding renal tissues in the TCGA cohort. B The schematic illustration of the CpG cg08521263 at the GABPA promoter. C The inverse correlation between the methylation of cg08521263 and GABPA mRNA levels in the TCGA cohort. D and E 5-AZA-treatment of A498 and 786-O cells upregulates GABPA expression coupled with the reduced methylation of cg08521263. Immunoblotting and pyrosequencing were used to assess GABPA expression and methylation, respectively. Three independent experiments were performed. F L-2-HG-treatment of A498 and 786-O cells inhibits GABPA expression. G The schematic illustration of conversion between L-2-HG and a-KG and related enzymes. H The genetic alterations in L2HGDH, MDH1/2, and LDHA/LDHB in tumors and from the TCGA ccRCC cohort and their correlation with GABPA expression. I The reduced 5-hmC accumulation in the GABPA sequence spanning cg08521263. Cells were treated with L-2-HG and ChIP assays were then carried out. J The restoration of L2HGDH expression upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. K MDH2 depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. L LDHB depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively
    Figure Legend Snippet: L-2-HG accumulation induces epigenetic silence of GABPA expression. A GABPA methylation is significantly higher in ccRCC tumors than in corresponding renal tissues in the TCGA cohort. B The schematic illustration of the CpG cg08521263 at the GABPA promoter. C The inverse correlation between the methylation of cg08521263 and GABPA mRNA levels in the TCGA cohort. D and E 5-AZA-treatment of A498 and 786-O cells upregulates GABPA expression coupled with the reduced methylation of cg08521263. Immunoblotting and pyrosequencing were used to assess GABPA expression and methylation, respectively. Three independent experiments were performed. F L-2-HG-treatment of A498 and 786-O cells inhibits GABPA expression. G The schematic illustration of conversion between L-2-HG and a-KG and related enzymes. H The genetic alterations in L2HGDH, MDH1/2, and LDHA/LDHB in tumors and from the TCGA ccRCC cohort and their correlation with GABPA expression. I The reduced 5-hmC accumulation in the GABPA sequence spanning cg08521263. Cells were treated with L-2-HG and ChIP assays were then carried out. J The restoration of L2HGDH expression upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. K MDH2 depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. L LDHB depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Techniques Used: Expressing, Methylation, Western Blot, Sequencing

    renal carcinoma cell line a498  (ATCC)


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    ATCC renal carcinoma cell line a498
    Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and <t>A498</t> (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.
    Renal Carcinoma Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Gut Microbiota-Derived Resveratrol Metabolites, Dihydroresveratrol and Lunularin, Significantly Contribute to the Biological Activities of Resveratrol"

    Article Title: Gut Microbiota-Derived Resveratrol Metabolites, Dihydroresveratrol and Lunularin, Significantly Contribute to the Biological Activities of Resveratrol

    Journal: Frontiers in Nutrition

    doi: 10.3389/fnut.2022.912591

    Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and A498 (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.
    Figure Legend Snippet: Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and A498 (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.

    Techniques Used:

    human renal epithelial cell line a498  (ATCC)


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    ATCC human renal epithelial cell line a498
    IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and <t>A498)</t> were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).
    Human Renal Epithelial Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Human Renal Fibroblasts, but Not Renal Epithelial Cells, Induce IL-1β Release during a Uropathogenic Escherichia coli Infection In Vitro"

    Article Title: Human Renal Fibroblasts, but Not Renal Epithelial Cells, Induce IL-1β Release during a Uropathogenic Escherichia coli Infection In Vitro

    Journal: Cells

    doi: 10.3390/cells10123522

    IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and A498) were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).
    Figure Legend Snippet: IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and A498) were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).

    Techniques Used: Infection

    a498 cell lines  (ATCC)


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    ATCC a498 cell lines
    A498 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    renal carcinoma a498 cell line  (ATCC)


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    ATCC renal carcinoma a498 cell line
    Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared <t>A498</t> MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.
    Renal Carcinoma A498 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/renal carcinoma a498 cell line/product/ATCC
    Average 99 stars, based on 1 article reviews
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    renal carcinoma a498 cell line - by Bioz Stars, 2024-05
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    1) Product Images from "Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids"

    Article Title: Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

    Journal: Scientific Reports

    doi: 10.1038/s41598-021-96288-6

    Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared A498 MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.
    Figure Legend Snippet: Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared A498 MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.

    Techniques Used: Staining, Confocal Microscopy, Fluorescence

    Correlation between cell cycle and integrated DAPI fluorescence in segmented nuclei. The cell cycle phases G1 (red), transition between G1/S (yellow) and G2/S ( a ) can be studied with the FUCCI construct which was transfected in A498 MCTSs (N = 40) ( b ). Based on integrated red and green intensity within each segmented nucleus, cells were classed as G1, G1/S or S/G2 positive ( c ). The FUCCI positive distribution of normalized integrated DAPI intensity per nucleus ( d ) could then be divided based on cell cycle position ( e – f ). Normalized DAPI intensity threshold against percentage of false positive rate (FPR) (red line), true positive S/G2 rate (TPR) (green line) and the difference between TPR and FPR (blue line) ( g ). The optimal threshold (1.25) is indicated at the TPR-FPR maximum (dashed orange line).
    Figure Legend Snippet: Correlation between cell cycle and integrated DAPI fluorescence in segmented nuclei. The cell cycle phases G1 (red), transition between G1/S (yellow) and G2/S ( a ) can be studied with the FUCCI construct which was transfected in A498 MCTSs (N = 40) ( b ). Based on integrated red and green intensity within each segmented nucleus, cells were classed as G1, G1/S or S/G2 positive ( c ). The FUCCI positive distribution of normalized integrated DAPI intensity per nucleus ( d ) could then be divided based on cell cycle position ( e – f ). Normalized DAPI intensity threshold against percentage of false positive rate (FPR) (red line), true positive S/G2 rate (TPR) (green line) and the difference between TPR and FPR (blue line) ( g ). The optimal threshold (1.25) is indicated at the TPR-FPR maximum (dashed orange line).

    Techniques Used: Fluorescence, Construct, Transfection

    Spatial distribution of S/G2 positive cells in A498 MCTSs cultured for 24- and 96-h. Normalized integrated DAPI intensity per nucleus data of segmented nuclei from A498 MCTSs cultured for 24 h (N = 10, orange) and 96 h (N = 10, blue) ( a ). The integrated intensity and volumetric parameters were used to classify G1 and S/G2 cells (percentages in each histogram). The surface of each MCTS was approximated and the distance between each nucleus and the MCTS perimeter was measured and placed in concentric layers automatically using MATLAB ( b ). The number of S/G2 positive cells ( c ) and fraction of S/G2 positive cells ( d ) was calculated for each concentric 10 µm wide layer, from 0 to 60 µm, for each MCTS. Bar plot distributions show mean and standard deviation for 24 (orange) and 96 (blue) hours old MCTSs. Scalebar is 20 µm.
    Figure Legend Snippet: Spatial distribution of S/G2 positive cells in A498 MCTSs cultured for 24- and 96-h. Normalized integrated DAPI intensity per nucleus data of segmented nuclei from A498 MCTSs cultured for 24 h (N = 10, orange) and 96 h (N = 10, blue) ( a ). The integrated intensity and volumetric parameters were used to classify G1 and S/G2 cells (percentages in each histogram). The surface of each MCTS was approximated and the distance between each nucleus and the MCTS perimeter was measured and placed in concentric layers automatically using MATLAB ( b ). The number of S/G2 positive cells ( c ) and fraction of S/G2 positive cells ( d ) was calculated for each concentric 10 µm wide layer, from 0 to 60 µm, for each MCTS. Bar plot distributions show mean and standard deviation for 24 (orange) and 96 (blue) hours old MCTSs. Scalebar is 20 µm.

    Techniques Used: Cell Culture, Standard Deviation

    Nucleus volume and local cell density with nucleus-MCTS perimeter distance as a continuous variable. Scatter plots shows the nucleus-MCTS perimeter distance, as a continuous variable ( a ), against nucleus volume ( b ) for A498 MCTSs cultured for 24 h (orange dots) and 96 h (blue dots). The alignment angle α, calculated as the angle between the MCTS center-nucleus center vector (r 1 ) and the plane spanned by the major (e 1 ) and intermediate (e 2 ) eigen vectors of the ellipsoid with the same second central momentum as the nucleus ( d ), is shown in the scatter plot against the nucleus-MCTS perimeter distance ( e ). The nuclear aspect ratio was studied by measuring the ratio between the major axis (p major ) and minor axis (p minor ) lengths ( g ) plotted against nucleus-MCTS perimeter distance ( h ). The nucleus-MCTS perimeter distance dependent trend in the scatter plots (red line with crosses) is calculated as the mean and median within 5 µm wide concentric layers for the nucleus volume, alignment angle and major/minor axes ratio respectively ( b , d , h ). Dotted line indicates the standard deviation and median absolute deviation. Box plot charts summarizes the data side by side for easier comparison ( c , f , i ). Significance was tested with two-way ANOVA followed by Tukey’s post-hoc multiple comparison (** p < 0.001). Scalebar is 20 µm.
    Figure Legend Snippet: Nucleus volume and local cell density with nucleus-MCTS perimeter distance as a continuous variable. Scatter plots shows the nucleus-MCTS perimeter distance, as a continuous variable ( a ), against nucleus volume ( b ) for A498 MCTSs cultured for 24 h (orange dots) and 96 h (blue dots). The alignment angle α, calculated as the angle between the MCTS center-nucleus center vector (r 1 ) and the plane spanned by the major (e 1 ) and intermediate (e 2 ) eigen vectors of the ellipsoid with the same second central momentum as the nucleus ( d ), is shown in the scatter plot against the nucleus-MCTS perimeter distance ( e ). The nuclear aspect ratio was studied by measuring the ratio between the major axis (p major ) and minor axis (p minor ) lengths ( g ) plotted against nucleus-MCTS perimeter distance ( h ). The nucleus-MCTS perimeter distance dependent trend in the scatter plots (red line with crosses) is calculated as the mean and median within 5 µm wide concentric layers for the nucleus volume, alignment angle and major/minor axes ratio respectively ( b , d , h ). Dotted line indicates the standard deviation and median absolute deviation. Box plot charts summarizes the data side by side for easier comparison ( c , f , i ). Significance was tested with two-way ANOVA followed by Tukey’s post-hoc multiple comparison (** p < 0.001). Scalebar is 20 µm.

    Techniques Used: Cell Culture, Plasmid Preparation, Standard Deviation

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    ATCC a498 cell lines
    A498 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC a498 human kidney carcinoma cell line
    Induction of cell death by EA in <t>A498</t> RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .
    A498 Human Kidney Carcinoma Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human kidney carcinoma cell line a498
    Induction of cell death by EA in <t>A498</t> RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .
    Human Kidney Carcinoma Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC kidney epithelial cell line a498
    Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line <t>A498.</t> The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.
    Kidney Epithelial Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC cell lines a498
    GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in <t>A498</t> and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively
    Cell Lines A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC renal carcinoma cell line a498
    Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and <t>A498</t> (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.
    Renal Carcinoma Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human renal epithelial cell line a498
    IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and <t>A498)</t> were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).
    Human Renal Epithelial Cell Line A498, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC renal carcinoma a498 cell line
    Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared <t>A498</t> MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.
    Renal Carcinoma A498 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Induction of cell death by EA in A498 RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: Induction of cell death by EA in A498 RCC cells. A498 cells were cells were treated with EA at 50 and 100 nM. Control cells received 0.1% DMSO (vehicle). All conditions were performed in triplicate. Cells were then incubated with additions for 24 or 48 h before measuring viability using the PrestoBlue ® assay (A) . A498 cells were treated with 100 nM EA or vehicle for 24 and 45 h durations. Apoptosis was determined by measuring cytoplasmic histone-associated-DNA-fragments using the Cell Death Detection ELISAPLUS assay kit (B) . A498 cells were treated with 100 nM EA or with 0.1% DMSO (control) for 24 and 46 h. Cells were then trypsinized, washed with ice cold PBS, and stained with Alexa Fluor ® 488 annexin V and PI and analyzed by flow cytometry (C) .

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Incubation, Prestoblue Assay, Staining, Flow Cytometry

    Caspases are not activated in-EA induced cell death. A498 cells cells were treated with 100 nM EA or 0.1% DMSO (control) for 43 h, or with 200 μM etoposide for 24 h. Cells were then harvested and stained with the FLICA reagent which only binds active caspases. Levels of active caspase were then determined by fluorescence (A) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-caspase-3 antibody. B-actin was probed as a control for protein loading (B) .

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: Caspases are not activated in-EA induced cell death. A498 cells cells were treated with 100 nM EA or 0.1% DMSO (control) for 43 h, or with 200 μM etoposide for 24 h. Cells were then harvested and stained with the FLICA reagent which only binds active caspases. Levels of active caspase were then determined by fluorescence (A) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-caspase-3 antibody. B-actin was probed as a control for protein loading (B) .

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Staining, Fluorescence, Western Blot

    EA induces autophagy in A498 cells. A498 cells were treated with 200 nM EA or 0.1% DMSO (control) for 46 h and with 500 nM rapamycin for 20 h. Autophagy was measured by staining autolysosomes and earlier autophagic compartments with the fluorescent probe Cyto-ID ® Green. Samples were then analyzed in the green (FL1) channel of the FACS Caliber flow cytometer (A) . Cells were treated with 200 nM EA or 0.1% DMSO (control) for 45 h and then stained with Hoechst nuclear stain and Cyto-ID ® Green detection reagent followed by fixing with 4% formaldehyde. The stained cells were then analyzed by fluorescence microscopy. Panels a and c show cells treated with 0.1% DMSO and panels b and d show cells treated with EA. Nuclei are stained in blue. Autolysosomes and earlier autophagic compartments are stained in green (B) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-LC3B antibody. B-actin was probed as a control for protein loading (C) .

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: EA induces autophagy in A498 cells. A498 cells were treated with 200 nM EA or 0.1% DMSO (control) for 46 h and with 500 nM rapamycin for 20 h. Autophagy was measured by staining autolysosomes and earlier autophagic compartments with the fluorescent probe Cyto-ID ® Green. Samples were then analyzed in the green (FL1) channel of the FACS Caliber flow cytometer (A) . Cells were treated with 200 nM EA or 0.1% DMSO (control) for 45 h and then stained with Hoechst nuclear stain and Cyto-ID ® Green detection reagent followed by fixing with 4% formaldehyde. The stained cells were then analyzed by fluorescence microscopy. Panels a and c show cells treated with 0.1% DMSO and panels b and d show cells treated with EA. Nuclei are stained in blue. Autolysosomes and earlier autophagic compartments are stained in green (B) . A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 48 h and protein was extracted. Western blot analysis was performed using an anti-LC3B antibody. B-actin was probed as a control for protein loading (C) .

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Staining, Flow Cytometry, Fluorescence, Microscopy, Western Blot

    Inhibition of autophagy does not affect EA-induced cell death. A498 cells were cultured in the presence and absence of 100 nM EA or, with EA and 1X NEAA in combination for 44 h. Autophagy was then determined by flow cytometry after staining with Cyto-ID ® (A) . A498 cells were treated with 150 nM EA, 0.1% DMSO, 1X NEAA, 200 μM VP16 or with 100 nM EA plus 1X NEAA for 46 h. Cell viability was then determined using the PrestoBlue ® assay (B) . A498 cells were treated as in (B) and then apoptosis was determined by measuring histone-associated DNA fragments by ELISA (C) .

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: Inhibition of autophagy does not affect EA-induced cell death. A498 cells were cultured in the presence and absence of 100 nM EA or, with EA and 1X NEAA in combination for 44 h. Autophagy was then determined by flow cytometry after staining with Cyto-ID ® (A) . A498 cells were treated with 150 nM EA, 0.1% DMSO, 1X NEAA, 200 μM VP16 or with 100 nM EA plus 1X NEAA for 46 h. Cell viability was then determined using the PrestoBlue ® assay (B) . A498 cells were treated as in (B) and then apoptosis was determined by measuring histone-associated DNA fragments by ELISA (C) .

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Inhibition, Cell Culture, Flow Cytometry, Staining, Prestoblue Assay, Enzyme-linked Immunosorbent Assay

    EA blocks the G 2 /M transition of the cell cycle. A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 45 h. The cells were then fixed and stained with PI. The PI content of cells was measured by flow cytometry as described under Methods.

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: EA blocks the G 2 /M transition of the cell cycle. A498 cells were treated with 200 nM EA or with 0.1% DMSO (control) for 45 h. The cells were then fixed and stained with PI. The PI content of cells was measured by flow cytometry as described under Methods.

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Staining, Flow Cytometry

    EA inhibits activation of AKT and ERK kinsases. A498 cells were cultured with 100 nM EA or with 0.1% DMSO (control) for the indicated times and protein was Isolated. Western blot analysis was performed as described under Methods using antibodies against AKT, ERK, and AMPK and their phosphorylated counterparts. B-actin was probed to control for protein loading. (+) control; Jurkat cell extract.

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy

    doi: 10.1186/1756-9966-32-57

    Figure Lengend Snippet: EA inhibits activation of AKT and ERK kinsases. A498 cells were cultured with 100 nM EA or with 0.1% DMSO (control) for the indicated times and protein was Isolated. Western blot analysis was performed as described under Methods using antibodies against AKT, ERK, and AMPK and their phosphorylated counterparts. B-actin was probed to control for protein loading. (+) control; Jurkat cell extract.

    Article Snippet: The A498 human kidney carcinoma cell line was purchased from ATCC and maintained in RPMI medium supplemented with 10% FBS and 100 units/ml penicillin/streptomycin (complete medium).

    Techniques: Activation Assay, Cell Culture, Isolation, Western Blot

    Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line A498. The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.

    Journal: PLoS ONE

    Article Title: Vitamin D Induction of the Human Antimicrobial Peptide Cathelicidin in the Urinary Bladder

    doi: 10.1371/journal.pone.0015580

    Figure Lengend Snippet: Expression of CYP27B1, VDR and CYP24A1 was detected on mRNA level in normal bladder cells (HBEP) and three bladder cells lines (TERT-NHUC, 5637 and T24) by RT-PCR ( A , C , E ) and on protein level in tissue from bladder biopsies by fluorescence immunohistochemistry ( B , D , F ). mRNA expression results were normalized to expression in the kidney epithelial cell line A498. The proteins are stained with AlexaFluor 594 (red), the cell nucleus with DAPI (blue). CYP27B1 protein was detected in the bladder mucosa where it was evenly distributed in the bladder epithelial layers with an abundance of staining seen in the peri-nuclear area, consistent with its association to mitochondria ( B ). VDR protein was localized in the cytoplasm and nucleus ( D ). Low levels of the CYP24A1 protein were detected in bladder mucosa ( F ). All images are captured at ×63 magnification and the right panels of images B, D and F show controls stained without primary antibodies against CYP27B1, VDR and CYP24A1, respectively.

    Article Snippet: Urinary bladder cell lines T24 (ATCC No. HTB-4) and 5637 (ATCC No. HTB-9) and kidney epithelial cell line A498 (ATCC No. HTB-44) were maintained in McCoy's medium, RPMI-1640 (Invitrogen) and Eagle's MEM (Sigma-Aldrich) respectively, supplemented with 10% fetal bovine serum (FBS).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Immunohistochemistry, Staining

    GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in A498 and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: GABPA-activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma

    doi: 10.1186/s13046-022-02382-6

    Figure Lengend Snippet: GABPA regulates proliferation, stemness and invasion of ccRCC cells. A GABPA-specific siRNAs and its expression vectors efficiently inhibit and overexpress GABPA in A498 and 786-O cells, respectively, as determined using immunoblotting. B GABPA depletion promotes while its overexpression inhibits cell proliferation. IncuCyte S3 Live-Cell Analysis System was employed to measure cell proliferation rate. C GABPA depletion increases while its overexpression reduces cells at S phase, as assessed using flow cytometry. D and E GABPA depletion increases while its overexpression reduces the stemness of A498 and 786-O cells. Scale bars: 200 µm. F and G GABPA depletion increases while its overexpression reduces the invasive capacity of A498 and 786-O cells. Scale bars: 100 µm. (H-J) GABPA regulates the expression of proliferation-, stemness- and EMT-related genes. (K) ccRCC stem cell (CD44 and CD105) and mesenchymal stem cell markers (CD90) are upregulated in GABPA-depleted cells. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Article Snippet: ccRCC-derived cell lines A498 (purchased from American Type Culture Collection, Manassas, VA) and 786-O cells (Shanghai Institute of Biochemistry and Cell Biology Cell Bank/Stem Cell Bank, Shanghai, China) were used in the present study.

    Techniques: Expressing, Western Blot, Over Expression, Flow Cytometry

    L-2-HG accumulation induces epigenetic silence of GABPA expression. A GABPA methylation is significantly higher in ccRCC tumors than in corresponding renal tissues in the TCGA cohort. B The schematic illustration of the CpG cg08521263 at the GABPA promoter. C The inverse correlation between the methylation of cg08521263 and GABPA mRNA levels in the TCGA cohort. D and E 5-AZA-treatment of A498 and 786-O cells upregulates GABPA expression coupled with the reduced methylation of cg08521263. Immunoblotting and pyrosequencing were used to assess GABPA expression and methylation, respectively. Three independent experiments were performed. F L-2-HG-treatment of A498 and 786-O cells inhibits GABPA expression. G The schematic illustration of conversion between L-2-HG and a-KG and related enzymes. H The genetic alterations in L2HGDH, MDH1/2, and LDHA/LDHB in tumors and from the TCGA ccRCC cohort and their correlation with GABPA expression. I The reduced 5-hmC accumulation in the GABPA sequence spanning cg08521263. Cells were treated with L-2-HG and ChIP assays were then carried out. J The restoration of L2HGDH expression upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. K MDH2 depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. L LDHB depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: GABPA-activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma

    doi: 10.1186/s13046-022-02382-6

    Figure Lengend Snippet: L-2-HG accumulation induces epigenetic silence of GABPA expression. A GABPA methylation is significantly higher in ccRCC tumors than in corresponding renal tissues in the TCGA cohort. B The schematic illustration of the CpG cg08521263 at the GABPA promoter. C The inverse correlation between the methylation of cg08521263 and GABPA mRNA levels in the TCGA cohort. D and E 5-AZA-treatment of A498 and 786-O cells upregulates GABPA expression coupled with the reduced methylation of cg08521263. Immunoblotting and pyrosequencing were used to assess GABPA expression and methylation, respectively. Three independent experiments were performed. F L-2-HG-treatment of A498 and 786-O cells inhibits GABPA expression. G The schematic illustration of conversion between L-2-HG and a-KG and related enzymes. H The genetic alterations in L2HGDH, MDH1/2, and LDHA/LDHB in tumors and from the TCGA ccRCC cohort and their correlation with GABPA expression. I The reduced 5-hmC accumulation in the GABPA sequence spanning cg08521263. Cells were treated with L-2-HG and ChIP assays were then carried out. J The restoration of L2HGDH expression upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. K MDH2 depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. L LDHB depletion upregulates GABPA expression coupled with increased 5-hmc and reduced methylation of cg08521263. All these effects are attenuated by addition of L-2-HG. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively

    Article Snippet: ccRCC-derived cell lines A498 (purchased from American Type Culture Collection, Manassas, VA) and 786-O cells (Shanghai Institute of Biochemistry and Cell Biology Cell Bank/Stem Cell Bank, Shanghai, China) were used in the present study.

    Techniques: Expressing, Methylation, Western Blot, Sequencing

    Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and A498 (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Nutrition

    Article Title: Gut Microbiota-Derived Resveratrol Metabolites, Dihydroresveratrol and Lunularin, Significantly Contribute to the Biological Activities of Resveratrol

    doi: 10.3389/fnut.2022.912591

    Figure Lengend Snippet: Anti-proliferative and anti-clonogenic effects of DHR and LUN in renal carcinoma cell lines at tissue-relevant concentrations. Anti-proliferative effects of DHR and LUN in 786-O (A) and A498 (B) renal carcinoma cells. Anti-clonogenic effects of DHR and LUN in 786-O (C) and A498 (D) cells. Three independent experiments were conducted. Data presented as mean ± SEM. Significant differences are indicated: * P < 0.05, ** P < 0.01.

    Article Snippet: Human colon adenocarcinoma cell line HT-29 (HTB-38), colorectal carcinoma cell line HCT-116 (CCL-247), renal carcinoma cell line A498 (HTB-44), and renal adenocarcinoma cell line 786-O (CRL-1932) were purchased from American Type Cell Collection (ATCC, Manassa, CA).

    Techniques:

    IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and A498) were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).

    Journal: Cells

    Article Title: Human Renal Fibroblasts, but Not Renal Epithelial Cells, Induce IL-1β Release during a Uropathogenic Escherichia coli Infection In Vitro

    doi: 10.3390/cells10123522

    Figure Lengend Snippet: IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and A498) were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).

    Article Snippet: Human renal medullary fibroblast cell line TK173 [ ] (kind gift from Professor Anton Jan van Zonneveld, Leiden University, Leiden, Netherlands), human renal epithelial cell line A498 (HTB-44, American Type Culture Collection (ATCC), Manassas, VA, USA) and the human immortalized proximal tubule epithelial cell line HK-2 (ATCC) were used in this study.

    Techniques: Infection

    Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared A498 MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.

    Journal: Scientific Reports

    Article Title: Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

    doi: 10.1038/s41598-021-96288-6

    Figure Lengend Snippet: Nuclear segmentation of confocal z-stacks. The full volume of DAPI stained and Histodenz cleared A498 MCTSs was acquired by confocal microscopy (3D rendering and optical section at z = 84 µm) ( a ). Outline of an in-house developed MATLAB script ( b ) used to segment all individual nuclei ( c ) for further volumetric and fluorescence intensity analysis. Scalebars are 20 µm.

    Article Snippet: The cells used in this study was renal carcinoma A498 cell line (ATCC© HTB-44™, ATCC) which was cultured in RPMI-1640 GlutaMAX (Thermo-Fischer Scientific) supplemented with 10% heat inactivated fetal bovine serum (Thermo-Fischer Scientific), 1 × MEM Non-essential amino acid solution and 25 mM HEPES (Thermo-Fischer Scientific).

    Techniques: Staining, Confocal Microscopy, Fluorescence

    Correlation between cell cycle and integrated DAPI fluorescence in segmented nuclei. The cell cycle phases G1 (red), transition between G1/S (yellow) and G2/S ( a ) can be studied with the FUCCI construct which was transfected in A498 MCTSs (N = 40) ( b ). Based on integrated red and green intensity within each segmented nucleus, cells were classed as G1, G1/S or S/G2 positive ( c ). The FUCCI positive distribution of normalized integrated DAPI intensity per nucleus ( d ) could then be divided based on cell cycle position ( e – f ). Normalized DAPI intensity threshold against percentage of false positive rate (FPR) (red line), true positive S/G2 rate (TPR) (green line) and the difference between TPR and FPR (blue line) ( g ). The optimal threshold (1.25) is indicated at the TPR-FPR maximum (dashed orange line).

    Journal: Scientific Reports

    Article Title: Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

    doi: 10.1038/s41598-021-96288-6

    Figure Lengend Snippet: Correlation between cell cycle and integrated DAPI fluorescence in segmented nuclei. The cell cycle phases G1 (red), transition between G1/S (yellow) and G2/S ( a ) can be studied with the FUCCI construct which was transfected in A498 MCTSs (N = 40) ( b ). Based on integrated red and green intensity within each segmented nucleus, cells were classed as G1, G1/S or S/G2 positive ( c ). The FUCCI positive distribution of normalized integrated DAPI intensity per nucleus ( d ) could then be divided based on cell cycle position ( e – f ). Normalized DAPI intensity threshold against percentage of false positive rate (FPR) (red line), true positive S/G2 rate (TPR) (green line) and the difference between TPR and FPR (blue line) ( g ). The optimal threshold (1.25) is indicated at the TPR-FPR maximum (dashed orange line).

    Article Snippet: The cells used in this study was renal carcinoma A498 cell line (ATCC© HTB-44™, ATCC) which was cultured in RPMI-1640 GlutaMAX (Thermo-Fischer Scientific) supplemented with 10% heat inactivated fetal bovine serum (Thermo-Fischer Scientific), 1 × MEM Non-essential amino acid solution and 25 mM HEPES (Thermo-Fischer Scientific).

    Techniques: Fluorescence, Construct, Transfection

    Spatial distribution of S/G2 positive cells in A498 MCTSs cultured for 24- and 96-h. Normalized integrated DAPI intensity per nucleus data of segmented nuclei from A498 MCTSs cultured for 24 h (N = 10, orange) and 96 h (N = 10, blue) ( a ). The integrated intensity and volumetric parameters were used to classify G1 and S/G2 cells (percentages in each histogram). The surface of each MCTS was approximated and the distance between each nucleus and the MCTS perimeter was measured and placed in concentric layers automatically using MATLAB ( b ). The number of S/G2 positive cells ( c ) and fraction of S/G2 positive cells ( d ) was calculated for each concentric 10 µm wide layer, from 0 to 60 µm, for each MCTS. Bar plot distributions show mean and standard deviation for 24 (orange) and 96 (blue) hours old MCTSs. Scalebar is 20 µm.

    Journal: Scientific Reports

    Article Title: Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

    doi: 10.1038/s41598-021-96288-6

    Figure Lengend Snippet: Spatial distribution of S/G2 positive cells in A498 MCTSs cultured for 24- and 96-h. Normalized integrated DAPI intensity per nucleus data of segmented nuclei from A498 MCTSs cultured for 24 h (N = 10, orange) and 96 h (N = 10, blue) ( a ). The integrated intensity and volumetric parameters were used to classify G1 and S/G2 cells (percentages in each histogram). The surface of each MCTS was approximated and the distance between each nucleus and the MCTS perimeter was measured and placed in concentric layers automatically using MATLAB ( b ). The number of S/G2 positive cells ( c ) and fraction of S/G2 positive cells ( d ) was calculated for each concentric 10 µm wide layer, from 0 to 60 µm, for each MCTS. Bar plot distributions show mean and standard deviation for 24 (orange) and 96 (blue) hours old MCTSs. Scalebar is 20 µm.

    Article Snippet: The cells used in this study was renal carcinoma A498 cell line (ATCC© HTB-44™, ATCC) which was cultured in RPMI-1640 GlutaMAX (Thermo-Fischer Scientific) supplemented with 10% heat inactivated fetal bovine serum (Thermo-Fischer Scientific), 1 × MEM Non-essential amino acid solution and 25 mM HEPES (Thermo-Fischer Scientific).

    Techniques: Cell Culture, Standard Deviation

    Nucleus volume and local cell density with nucleus-MCTS perimeter distance as a continuous variable. Scatter plots shows the nucleus-MCTS perimeter distance, as a continuous variable ( a ), against nucleus volume ( b ) for A498 MCTSs cultured for 24 h (orange dots) and 96 h (blue dots). The alignment angle α, calculated as the angle between the MCTS center-nucleus center vector (r 1 ) and the plane spanned by the major (e 1 ) and intermediate (e 2 ) eigen vectors of the ellipsoid with the same second central momentum as the nucleus ( d ), is shown in the scatter plot against the nucleus-MCTS perimeter distance ( e ). The nuclear aspect ratio was studied by measuring the ratio between the major axis (p major ) and minor axis (p minor ) lengths ( g ) plotted against nucleus-MCTS perimeter distance ( h ). The nucleus-MCTS perimeter distance dependent trend in the scatter plots (red line with crosses) is calculated as the mean and median within 5 µm wide concentric layers for the nucleus volume, alignment angle and major/minor axes ratio respectively ( b , d , h ). Dotted line indicates the standard deviation and median absolute deviation. Box plot charts summarizes the data side by side for easier comparison ( c , f , i ). Significance was tested with two-way ANOVA followed by Tukey’s post-hoc multiple comparison (** p < 0.001). Scalebar is 20 µm.

    Journal: Scientific Reports

    Article Title: Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

    doi: 10.1038/s41598-021-96288-6

    Figure Lengend Snippet: Nucleus volume and local cell density with nucleus-MCTS perimeter distance as a continuous variable. Scatter plots shows the nucleus-MCTS perimeter distance, as a continuous variable ( a ), against nucleus volume ( b ) for A498 MCTSs cultured for 24 h (orange dots) and 96 h (blue dots). The alignment angle α, calculated as the angle between the MCTS center-nucleus center vector (r 1 ) and the plane spanned by the major (e 1 ) and intermediate (e 2 ) eigen vectors of the ellipsoid with the same second central momentum as the nucleus ( d ), is shown in the scatter plot against the nucleus-MCTS perimeter distance ( e ). The nuclear aspect ratio was studied by measuring the ratio between the major axis (p major ) and minor axis (p minor ) lengths ( g ) plotted against nucleus-MCTS perimeter distance ( h ). The nucleus-MCTS perimeter distance dependent trend in the scatter plots (red line with crosses) is calculated as the mean and median within 5 µm wide concentric layers for the nucleus volume, alignment angle and major/minor axes ratio respectively ( b , d , h ). Dotted line indicates the standard deviation and median absolute deviation. Box plot charts summarizes the data side by side for easier comparison ( c , f , i ). Significance was tested with two-way ANOVA followed by Tukey’s post-hoc multiple comparison (** p < 0.001). Scalebar is 20 µm.

    Article Snippet: The cells used in this study was renal carcinoma A498 cell line (ATCC© HTB-44™, ATCC) which was cultured in RPMI-1640 GlutaMAX (Thermo-Fischer Scientific) supplemented with 10% heat inactivated fetal bovine serum (Thermo-Fischer Scientific), 1 × MEM Non-essential amino acid solution and 25 mM HEPES (Thermo-Fischer Scientific).

    Techniques: Cell Culture, Plasmid Preparation, Standard Deviation