caki 1 mccoy human renal cell carcinoma lines  (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: Representative images from the wound closure assay. ( A ) 786-O cells were imaged at 0, 10, and 20 h. ( B ) Caki-1 cells were imaged at 0, 24, and 48 h. The compounds are arranged from top to bottom: DMSO, KC7F2, GN44028, FM19G11, and sunitinib. Scale bar: 100 µm. Magnification 400×.

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: Wound Closure Assay

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: Quantitative analysis of wound closure dynamics. ( A ) 786-O cells (0–20 h). ( B ) Caki-1 cells (0–48 h). Y-axis: Wound closure in culture [%]. The compounds are arranged from top to bottom: DMSO (yellow), KC7F2 (blue), GN44028 (green), FM19G11 (red), and sunitinib (black). Data are presented as mean ± SD from independent biological experiments, each measured in technical replicates ( n = 18–23). **** p < 0.0001 vs. DMSO (one-way ANOVA + 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:

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: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Identification and validation of CuCOX peak in SEC-ICP-MS chromatogram. A , Top : representative SEC-ICP-MS chromatograms of lysates from 786-O and RCC4 cells grown in 0.14 μM or 30 μM Cu for 48 h. Bottom: UV-Vis absorbance isoplots collected throughout the entire chromatography run. The legend shows the logarithmic scale for absorbance in the isoplot. Note that, because absorbance was measured before the ICP-MS detector, there is a slight shift in the timeline between the chromatogram and the spectra. B , measurement of absorbance from the 5 min fraction presented on a linear scale shows Cu-induced peaks at 420 and 600 nm. C , box whisker plots show relative abundance of Cu measured by SEC-ICP-MS for CuCOX and MTs, and by ICP-MS for total Cu content in 786-O (3 biological and 2 technical replicates) and RCC4 (4 biological and 2 technical replicates) cells. D , Western blot of indicated proteins eluted from the CuCOX peak and the inputs are shown below. E , oxygen consumption rate (OCR) from representative seahorse mitochondrial stress tests in 786-O and RCC4 cells (3 biological replicates). Quantification of basal and maximal respiration (post-FCCP injection), and respiration coupled to ATP production. Means ± SD are shown. p -values for basal respiration in 786-O cells was calculated using a two-tailed, paired t test. F , SEC-ICP-MS chromatograms, quantification of CuCOX and MT peaks, total Cu level, and Seahorse measurements of OCR for RPTEC TH1 and TERT1 cell lines (3 biological replicates and 2 technical replicates). Box-and-whisker plots display median, minimum and maximum values and all individual data points. Unless otherwise indicated, p -values were determined using a two-tailed, unpaired t test. All Cu measurements are normalized to total phosphorus (P). HMW, high molecular weight fraction; LMW, low molecular weight fraction; MT, metallothioneins

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Biomarker Discovery, Chromatography, Whisker Assay, Western Blot, Injection, Two Tailed Test, High Molecular Weight, Molecular Weight

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Galactose induces oxidative phosphorylation and increases Cu content and allocation to CuCOX in 786-O RCC cells. A , schematic representation of the Leloir pathway converting galactose into glucose. B , OCR and mitochondrial ATP production in response to 48 h treatment with galactose ( red ) as compared to glucose ( black ) (3 biological replicates). C , SEC-ICP-MS chromatograms of Cu ( top ) and corresponding UV-Vis absorbance plots in lysates from 786-O cells treated with 10 mM glucose or 10 mM galactose for 48 h. The legend shows the logarithmic scale for absorbance in the isoplot. D , box whiskers show quantification of Cu allocated to CuCOX and MTs, and total Cu content in lysates from 786-O cells grown in glucose- or galactose-containing media (7 biological replicates with 2 technical replicates for each). E , Western blot of total input lysates and CuCOX eluates from 786-O grown in glucose or galactose media. F , Schematic model of mitochondrial electron transport chain along with localization of inhibitors for each complex. G , quantification of SEC-ICP-MS peaks corresponding to Cu in CuCOX and MTs, and total Cu in lysates of 786-O cells treated with media containing glucose, galactose, or galactose with the indicated electron transport chain inhibitors added for 3 h before collection. ROT, rotenone (200 nM, three biological replicates in technical duplicates), IACS, IACS-10759 (1 nM, 3biological replicates in technical duplicates), MET, metformin (25 mM, 3 biological replicates in technical duplicates), AM, antimycin A (1 nM, 4 biological replicates in technical duplicates), MYX, myxothiazol (1 nM, 4 biological replicates run in technical duplicates), KCN, potassium cyanide (0.5 mM, 4 biological replicates in technical duplicates), NaN3, sodium azide (0.5 mM, 4 biological replicates in technical duplicates). Box-and-whisker plots displayed the median, minimum and maximum values, and all individual data points. All p -values were calculated using a two-tailed, unpaired t test and compared the treatment groups with the galactose-only condition.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Phospho-proteomics, Western Blot, Whisker Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Kinetics of allocation of 63Cu tracer into CuCOX. A , schematic representation of tracing 63 Cu. B , timeline of 63 Cu tracking into CuCOX. C and D , SEC-ICP-MS chromatograms of exogenous 63 Cu in lysates from 786-O and RCC4 cells at the indicated time points. E and F , quantification by SEC-ICP-MS of exogenous 63 Cu in CuCOX, MTs and total exogenous 63 Cu in 786-O and RCC4 cells. Cu abundance was normalized to Phosphorus (P). In all experiments: n = 4, SEM ± SD are shown; p- values were calculated using a two-tailed, unpaired t test and indicate significance of the difference between 10 or 30 μM Cu compared to 0.14 μM Cu. Experiments were performed in 3 technical replicates twice and a representative experiment is shown.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Allocation of tracer 63 Cu to CuCOX in 786-O and RCC4 cells chronically grown in low or high Cu media. A , timeline of the experiment: AC, exposure to 30 μM 63 Cu for 48 h of cells grown continuously in low Cu media; CH, exposure to 30 μM 63 Cu for 48 h of cells grown in high Cu media. B , representative examples of Cu SEC-ICP-MS chromatograms from cells grown in Cu low media and exposed to 30 μM 63 Cu for 48 h (AC) or cells grown continuously in media containing 30 μM Cu and then treated with the same concentration of 63 Cu (CH). C , quantification of 63 Cu in total exogenous and endogenous Cu measured by SEC-ICP-MS. D , quantification of 63 Cu in exogenous and endogenous CuCOX peak from SEC-ICP-MS chromatograms. E , representative examples of Zn SEC-ICP-MS chromatograms as described in B. F , quantification of total cellular Zn based on SEC-ICP-MS. G , representative examples of Fe SEC-ICP-MS chromatograms as described in B . H , quantification of total cellular Fe based on SEC-ICP-MS chromatograms. Metals’ abundance was normalized to Phosphorus (P). In all experiments: Means ± SD are shown; n = 4 biological replicates; p - values were calculated using a two-tailed, unpaired t test.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Concentration Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Contribution of Cu transporters to the allocation of tracer 63 Cu to CuCOX. A , timeline of experiments for cells cultured under low or high Cu conditions and subsequently exposed to 30 μM 63 Cu for 48 h (AC and CH, respectively) or to 0.14 μM 63 Cu for 48 h (LO). B–E , effects of the knockdowns of indicated transporters on the allocation of exogenous 63 Cu and total content of Zn and Fe measured by SEC-ICP-MS in AC conditions for 786-O ( top ) and RCC4 ( bottom ) cells. F-I , effects of the knockdowns of indicated transporters on the allocation of exogenous 63 Cu and total content of Zn and Fe measured by SEC-ICP-MS in CH conditions for both cell lines. J and K , Western blots showing efficiency of the indicated knockdowns for AC ( J ) and CH ( K ) conditions. L and M , RT-PCR validation of CTR1 KD under AC ( L ) and CH ( M ) conditions. NT-non-targeting siRNA. Metals’ abundance was normalized to Phosphorus (P). N , Western blots showing expression of ZNT1 in RCC4 and 786-O cells and the effectiveness of the knockout. O , effects of the ZNT1 knockout on the total exogenous 63 Cu uptake and 63 Cu allocation to CuCOX, as well as total content of Zn, and Fe measured by SEC-ICP-MS in AC conditions for RCC4 cells. P , SEC-ICP-MS chromatograms of lysates from 786-O cells grown in 0.14 μM Cu, treated with non-targeting (NT) or CTR1 siRNAs, and exposed to 0.14 μM 63 Cu for 48 h. Q , quantification of total 63 Cu, and 63 Cu allocated to CuCOX and MT measured by SEC-ICP-MS in response to CTR1 KD. R , DepMap gene-effect scores distribution and median across all RCC cell lines for five metal transporter genes. A blue dot marks 786-O cell line. Biological replicates: B–I and Q , n = 4 (except for NT treatment in RCC4 in B–E and CTR1 KD in 786-O in F–I where n = 3); L and M , n = 3; O, n = 6. Means ± SD are shown; p - values were calculated using a two-tailed, unpaired t test and comparing the significance of changes caused by specific si/sgRNA to NT si/sgRNA.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Expressing, Knock-Out, Two Tailed Test

Journal: STAR Protocols

Article Title: Protocol to generate human stem cell-derived CD70-directed allogeneic CAR-NKT cells for treating renal cell carcinoma

doi: 10.1016/j.xpro.2025.104340

Figure Lengend Snippet: Evaluation of the in vitro antitumor efficacy of Allo CAR70-NKT cells (A) Experimental design to study the in vitro antitumor efficacy of Allo CAR70-NKT cells against human RCC cell lines. (B) Schematics showing the indicated human tumor cell lines. (C) Tumor cell killing data at 24 h (n = 4). (D) ELISA analyses of IFN-γ and TNF-α production by Allo CAR70-NKT cells after 24-hour coculture with 786-O-FG cells (n = 4). (E) Experimental design to Study the tumor cell killing mechanisms of Allo CAR70-NKT cells mediated by NKRs. (F) Tumor cell killing data at 24 h (E:T ratio = 0.5:1; n = 4). Representative of 3 experiments. Data are presented as the mean ± SEM. ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 by Student’s t test (D) or one-way ANOVA (C and F).

Article Snippet: Human renal cell carcinoma cell line 786-O , ATCC , CRL-1932.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay