human papillomavirus transformed kidney cell line hk  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Exploratory Multi-Level Analysis of the HIF Axis in Clear-Cell Renal Cell Carcinoma and Evaluation of GN44028 as an Experimental HIF Pathway-Modulating Compound

doi: 10.3390/ijms27083505

Figure Lengend Snippet: Cytotoxicity of HIF inhibitors and sunitinib in renal cell lines: ( A ) 786-O, ( B ) Caki-1, and ( C ) HK-2. Dose–response curves (SRB, 72 h of incubation). Data are presented as mean ± SD from independent biological experiments, each measured in technical replicates ( n = 6). Curves were fitted using four-parameter logistic regression.

Article Snippet: 786-O (cRL1932TM), Caki-1 (McCoy) human renal cell carcinoma lines, and HK-2 (cRL2190TM) human proximal kidney tubule-derived cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: Exploratory Multi-Level Analysis of the HIF Axis in Clear-Cell Renal Cell Carcinoma and Evaluation of GN44028 as an Experimental HIF Pathway-Modulating Compound

doi: 10.3390/ijms27083505

Figure Lengend Snippet: Cell cycle analysis by propidium iodide staining and flow cytometry in 786-O ( A ), Caki-1 ( B ), and HK-2 ( C ) cells treated with KC7F2, FM19G11, GN44028, and sunitinib at IC 50 concentrations for 24 h. Bar graphs show mean ± SD from independent biological experiments, each measured in technical replicates ( n = 3–19). GN44028 was associated with an increased sub-G1 fraction in ccRCC cell lines and G2/M arrest in HK-2 cells. Sunitinib induces G0/G1 arrest in 786-O cells. **** p < 0.0001, * p < 0.05 vs. DMSO (ANOVA with Dunnett’s test).

Article Snippet: 786-O (cRL1932TM), Caki-1 (McCoy) human renal cell carcinoma lines, and HK-2 (cRL2190TM) human proximal kidney tubule-derived cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Cell Cycle Assay, Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Exploratory Multi-Level Analysis of the HIF Axis in Clear-Cell Renal Cell Carcinoma and Evaluation of GN44028 as an Experimental HIF Pathway-Modulating Compound

doi: 10.3390/ijms27083505

Figure Lengend Snippet: Heatmap of transcriptional responses of hypoxia- and HIF-regulated genes in renal cell carcinoma cell lines following pharmacological inhibition of the HIF pathway. The heatmap shows log 2 (fold change) values of mRNA expression relative to vehicle-treated controls (DMSO) for 786-O and Caki-1 cells after 72 h of treatment with KC7F2, GN44028, FM19G11, or sunitinib (SUN) at IC 50 concentrations. Normal proximal tubule cells (HK-2) were excluded from visualisation. All qPCR data are presented as log 2 fold change relative to the vehicle-treated controls (DMSO). Red, green, and white colours indicate downregulation, upregulation, and no change in expression, respectively. The colour scale represents log 2 FC values ranging from −5 to +5. Genes were hierarchically clustered (average linkage, Euclidean distance), and treatment conditions were ordered manually. Each value represents the mean of two independent biological experiments, each with three technical replicates.

Article Snippet: 786-O (cRL1932TM), Caki-1 (McCoy) human renal cell carcinoma lines, and HK-2 (cRL2190TM) human proximal kidney tubule-derived cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Inhibition, Expressing

Journal: bioRxiv

Article Title: Empagliflozin targets a renal neuro-epithelial-immune axis in heart failure

doi: 10.64898/2026.03.31.715595

Figure Lengend Snippet: (A) Urinary norepinephrine (NE) excretion at randomization and posttreatment (n = 8-9 rats per group). (B) Renal cortical norepinephrine (NE) concentration. (C) Sodium-glucose cotransporter 2 (SGLT2; Slc5a2 ) mRNA expression in HK-2 human proximal tubule cells treated with vehicle, norepinephrine (NE), empagliflozin (EMPA), or NE + EMPA (n = 6). Data are presented as mean ± SEM. For in vivo experiments, statistical analysis was performed using an unpaired Student’s t test for randomization data (panel A, n = 17 rats per group) and two-way ANOVA followed by Tukey’s post hoc test (panels A and B). For in vitro experiments, one-way ANOVA followed by Tukey’s post hoc test was used. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: The human kidney cell line (HK-2) was purchased from ATCC (CRL-2190) and cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO Invitrogen), 1% penicillin/streptomycin (GIBCO Invitrogen), 50 ng/ml bovine pituitary extract (BPE) (13028-014, Thermo Fisher Scientific, Waltham, MA), and 5 ng/ml human recombinant epidermal growth factor (hEGF) (10450-013, Thermo Fisher Scientific) at 37°C, 5% CO 2 in a humidified tissue culture incubator.

Techniques: Concentration Assay, Expressing, In Vivo, In Vitro

Journal: bioRxiv

Article Title: Empagliflozin targets a renal neuro-epithelial-immune axis in heart failure

doi: 10.64898/2026.03.31.715595

Figure Lengend Snippet: (A) IL-6 mRNA expression in HK-2 cells treated with vehicle (Control), empagliflozin (EMPA, 1 μM), norepinephrine (NE, 1 μM), or combined NE + EMPA. IL-6 mRNA levels were normalized to cyclophilin and expressed as a percentage of control. Each data point represents five independent experiments performed in duplicate. (B) IL-6 concentration in the supernatant of HK-2 cells under the same experimental conditions, measured by ELISA and normalized to protein content. Individual data points represent nine independent experiments performed in duplicate. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ***P < 0.001 and ****P < 0.0001.

Article Snippet: The human kidney cell line (HK-2) was purchased from ATCC (CRL-2190) and cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO Invitrogen), 1% penicillin/streptomycin (GIBCO Invitrogen), 50 ng/ml bovine pituitary extract (BPE) (13028-014, Thermo Fisher Scientific, Waltham, MA), and 5 ng/ml human recombinant epidermal growth factor (hEGF) (10450-013, Thermo Fisher Scientific) at 37°C, 5% CO 2 in a humidified tissue culture incubator.

Techniques: Expressing, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Calcified Tissue International

Article Title: Bone from Healthy Individuals and Patients with CKD Expresses the Sodium-Glucose Co-transporter-2 (SGLT2)

doi: 10.1007/s00223-026-01498-7

Figure Lengend Snippet: Preliminary qPCR amplification for primer selection. A and B Unsuccessful preliminary qPCR amplification using 267 and 430 bp primer pairs for SLC5A2 for MG-63 cells, respectively. The amplification plot displays the performance designed to amplify as a product of the human SLC5A2 transcript ( NM_003041.4 ), with its specific binding sites shown below. While the primers yielded a clear amplification signal in the positive control HK-2 renal cells, no detectable expression was observed in the osteoblast-like MG-63 cell line. Due to the lack of amplification in MG-63 cells, these primer sets were deemed unsuitable for the study. ( C ) Successful validation of the selected 108 bp primer for SLC5A2. This primer pair shows robust and comparable amplification in both MG-63 and HK-2 cell lines between 22 and 24 cycles, confirming its suitability for subsequent comparative gene expression analysis by qPCR

Article Snippet: Human osteoblast-like (MG-63) and human kidney (HK-2) cell lines were obtained from the American Type Culture Collection (ATCC CRL-1427 and -2190, USA).

Techniques: Amplification, Selection, Binding Assay, Positive Control, Expressing, Biomarker Discovery, Gene Expression

Journal: Calcified Tissue International

Article Title: Bone from Healthy Individuals and Patients with CKD Expresses the Sodium-Glucose Co-transporter-2 (SGLT2)

doi: 10.1007/s00223-026-01498-7

Figure Lengend Snippet: Relative quantification of SLC5A2 gene expression. A Raw Ct values of two biologicals (in duplicates) showing comparable SLC5A2 expression in MG-63 and HK-2 cells; for the first time, the SLC5A2 expression in osteoblast-like MG-63 cells is confirmed; B Raw Ct values of three healthy individuals bone samples and ten bone samples from patients with CKD (in duplicates) indicating that the SLC5A2 expression is lower in bone samples from patients with CKD. After ΔΔCt normalization by GAPDH, no significant difference is seen between C cell lines (two biologicals in duplicates), or between D bone samples (mean of duplicates) from healthy subjects and patients with CKD. GAPDH variation in expression (above 3.5 Cts) prevented a suitable normalization of SLC5A2 expression in healthy subjects versus patients with CKD; thus, each condition was normalized to its own GAPDH expression for representation purposes

Article Snippet: Human osteoblast-like (MG-63) and human kidney (HK-2) cell lines were obtained from the American Type Culture Collection (ATCC CRL-1427 and -2190, USA).

Techniques: Quantitative Proteomics, Gene Expression, Expressing

Journal: Calcified Tissue International

Article Title: Bone from Healthy Individuals and Patients with CKD Expresses the Sodium-Glucose Co-transporter-2 (SGLT2)

doi: 10.1007/s00223-026-01498-7

Figure Lengend Snippet: Absolute quantification of SLC5A2 and GAPDH transcript copy numbers in cells (two biologicals in duplicates) and human bone (mean of duplicates from three healthy individuals’ bone and ten samples from patients with CKD) after 40 cycles. The use of standard curves and linear regressions using dilutions in log 10 -3, -4, -5, and -6 from experimental controls (HK-2 cells and apparently healthy individuals´ bone samples) templates allowed the estimation of an unknown number of copies for each primer individually. This analysis confirms comparable absolute SLC5A2 transcript levels between HK-2 and osteoblast-like MG-63 cells, while GAPDH levels were modestly lower in MG-63 cells. In human bone tissue, a non-significant trend towards higher SLC5A2 copies was observed in samples from patients with chronic kidney disease (CKD) compared to samples from healthy subjects. Critically, GAPDH copy numbers were significantly reduced in CKD bone, demonstrating its instability as a reference gene in this pathology and reinforcing the value of absolute quantification for accurate expression analysis while analyzing bone samples. Additionally, internal controls also using pools from biologicals of HK-2 cells and healthy bone (red dashed line indicated within each ‘y’ axis) were adopted as an extra sample (in duplicates) to corroborate the unknown individual sample results; this reference obtained its number of copies within the range of their tested biologicals

Article Snippet: Human osteoblast-like (MG-63) and human kidney (HK-2) cell lines were obtained from the American Type Culture Collection (ATCC CRL-1427 and -2190, USA).

Techniques: Quantitative Proteomics, Expressing