murine renal cortical adenocarcinoma cell line renca  (ATCC)

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted ncSTING ADCs drive antitumor activity across multiple tumor models with multiple tumor antigens. Mean tumor volume over time of ( A ) human CD228-LL2 tumor–bearing C57BL/6 mice, ( B ) murine B7-H4-EMT6 tumor–bearing C57BL/6 mice, ( C ) murine B7-H4-Renca tumor–bearing Balb/c mice, and ( D ) murine IB6-CT26 tumor–bearing Balb/c mice following treatment with three weekly intraperitoneal doses (arrows) of the indicated ADCs. Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Activity Assay, In Vivo

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted STING agonist delivery via an ADC minimizes systemic cytokine induction compared with systemic delivery of the released payload. A and C, Mean tumor volume over time of Renca tumor–bearing Balb/c mice following treatment with three weekly intravenous ( A ) or intraperitoneal ( C ) doses (arrows) of the indicated ADCs or released payload. B and D, Quantification of IL-6 in the plasma 3, 6, and 24 hours following treatment as in A and C . Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare AUC.3 values ( A and C ) or cytokine levels ( B and D ) for each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Clinical Proteomics, In Vivo

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted ncSTING ADCs with a WT Fc backbone drive enhanced antitumor activity in some, but not all, tumor models. Mean tumor volume over time of ( A ) human CD228-LL2 tumor–bearing C57BL/6 mice, ( B ) murine B7-H4-EMT6 tumor–bearing C57BL/6 mice, ( C ) murine B7-H4-Renca tumor–bearing Balb/c mice, ( D ) murine IB6-CT26 tumor–bearing Balb/c mice, ( E ) MC38 tumor–bearing C57BL/6 mice, and ( F ) MC38 tumor–bearing C57BL/6 WT and STING-deficient mice following treatment with the three weekly intraperitoneal doses (arrows) or a single dose (denoted by “x1”) of the indicated ADCs. Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare AUC.3 values for each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Activity Assay, In Vivo

Journal: The Journal of Biological Chemistry

Article Title: Small molecule intervention of actin-binding protein profilin1 reduces tumor angiogenesis in renal cell carcinoma

doi: 10.1016/j.jbc.2025.111033

Figure Lengend Snippet: Demonstration of C74’s ability to diminish tumor angiogenesis . A , chemical structure of C74. B - C , representative images ( panel B ; 4 × magnification) and quantification ( panel C ; based on 10 × field images) of CD31 + cells in subcutaneously established RENCA tumors subjected to daily direct intratumoral injection for 19 days with either C74 or equivalent amount of DMSO control in BALB/c mice (two-tailed Student’s t test, ∗∗, p < 0.01; n = 5 mice/group; the scale bar represents 200 μm). DMSO, dimethyl sulfoxide.

Article Snippet: RENCA renal cell carcinoma (RCC) cells (American Type Culture Collection [ATCC], CRL-2947) were cultured in RPMI supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin.

Techniques: Injection, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Small molecule intervention of actin-binding protein profilin1 reduces tumor angiogenesis in renal cell carcinoma

doi: 10.1016/j.jbc.2025.111033

Figure Lengend Snippet: Feasibility of ultrasound-guided tumor-localized delivery of Pfn1 inhibitor to achieve antiangiogenic and therapeutic benefit. A , experimental schema of tumor microbubble studies (MB–microbubbles). B , ultrasound images ( panel B ) showing MB infiltration into tumor vasculature ( white arrows ), followed by UTMD and no contrast remaining in the tumor space. C – J , results of C74-MB ( panels C through F ) and UP6-MB ( panels G – J ) tumor model studies. Images of subcutaneous RENCA tumors subjected to UTMD of indicated MBs ( panels C and G ) with end point tumor burden comparison between the control and experimental treatment groups ( panels D and H ). Representative H&E images of tumor histosections ( panels E and I; white arrows indicate functional tumor-associated blood vessels with presence of red blood cells) and the associated quantification ( panels F and I ) of the average number of tumor-associated blood vessels per 20X field of observation between the two treatment groups (Mann-Whitney test, ∗: p < 0.05; ∗∗ p < 0.01, ∗∗∗: p < 0.001; data pooled from 8-9 mice per group; the scale bar represents 50 μm). Pfn1, profilin1; UTMD, ultrasound-targeted microbubble destruction.

Article Snippet: RENCA renal cell carcinoma (RCC) cells (American Type Culture Collection [ATCC], CRL-2947) were cultured in RPMI supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin.

Techniques: Comparison, Control, Functional Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: A strategically designed homogeneous antibody-drug conjugate improves safety and therapeutic index in renal cell carcinoma

doi: 10.64898/2026.01.12.699160

Figure Lengend Snippet: ( a ) Schematic representation of the synthesis of an antibody fluorophore conjugate (AFC) by using the MTGase reaction as described for the ADC. First, the native monoclonal antibody was deglycosylated in the presence of the PNGase F enzyme, and then the Cy5-fluorophore-linker was attached to the antibody at Q295 in the presence of the MTGase. ( b ) SDS-PAGE (fluorescence imaging and Coomassie staining) of KIM-1 antibody, deglycosylated antibody, LT-025, and Cy5-AFC. The data shows attachment of the Cy5 fluorophore in the heavy chain of the antibody. No conjugation of Cy5 was observed in the light chain. ( c ) Schematic representation of the cellular internalization of the ADC and AFC. The AFC first binds with the KIM-1 receptor on the cell surface, and it gets endocytosed into the cell and is transported to the lysosomal compartment. ( d ) Internalization study of Cy5 AFC at different time points. 50 μg/ml of AFC. An increase in MFI (Cy5) over time means increased binding/internalization of AFC onto the Renca RCC cell line. ( e ) Confocal microscopy images showing the internalization of the AFC in Renca cells. The images show successful cellular internalization of AFC (50 μg/ml) within 0.5 h and 4 h. An increased cellular internalization and colocalization at the lysosomal compartment were observed in 4 h. Scale bar = 5 μm.

Article Snippet: Renca cells (1×10 6 cells in PBS) were injected subcutaneously into flank region of 8-10 weeks syngeneic male balb/c mice (Charles River Laboratories, Strain Code 028).

Techniques: SDS Page, Fluorescence, Imaging, Staining, Conjugation Assay, Binding Assay, Confocal Microscopy

Journal: bioRxiv

Article Title: A strategically designed homogeneous antibody-drug conjugate improves safety and therapeutic index in renal cell carcinoma

doi: 10.64898/2026.01.12.699160

Figure Lengend Snippet: Characterization of transduction of KIM-1 on RENCA cells using ( a ) Western blotting and ( b ) flow cytometry. The mean fluorescence intensity (MFI) was plotted in the bar graph. FITC conjugated anti-mouse KIM-1 antibody was used to probe the KIM-1 expression ( c ) The KIM-1 AFC internalization in Normal Renca and Renca KIM-1++ by flow cytometry. The histogram shows a notable increase in the cellular internalization both at 1 and 4 h. ( d ) The bar graph represents the MFI of AFC internalization in Renca and Renca KIM-1++ by flow cytometry. The data show a significant increase in cellular internalization. The data were represented as mean ± SEM (n = 3, two-way ANOVA followed by Tukey’s multiple comparison test).

Article Snippet: Renca cells (1×10 6 cells in PBS) were injected subcutaneously into flank region of 8-10 weeks syngeneic male balb/c mice (Charles River Laboratories, Strain Code 028).

Techniques: Transduction, Western Blot, Flow Cytometry, Fluorescence, Expressing, Comparison

Journal: bioRxiv

Article Title: A strategically designed homogeneous antibody-drug conjugate improves safety and therapeutic index in renal cell carcinoma

doi: 10.64898/2026.01.12.699160

Figure Lengend Snippet: Quantification of the colocalization between AFC (green) signal with Lysotracker (red) signal in Renca cells at 0.5 and 4 h. The data represent significantly higher colocalization at 4 h. The data were represented as mean ± SEM (n = 5, Unpaired t -test with Welch’s correction).

Article Snippet: Renca cells (1×10 6 cells in PBS) were injected subcutaneously into flank region of 8-10 weeks syngeneic male balb/c mice (Charles River Laboratories, Strain Code 028).

Techniques:

Journal: bioRxiv

Article Title: A strategically designed homogeneous antibody-drug conjugate improves safety and therapeutic index in renal cell carcinoma

doi: 10.64898/2026.01.12.699160

Figure Lengend Snippet: Cell viability and IC 50 of ADC on ( a ) normal Renca and ( b ) Renca KIM-1++, obtained by MTT assay. The LT-025 exhibits significant cell death with the IC 50 values in the nanomolar range. A lower IC 50 value was observed in Renca KIM-1++ than in normal Renca because of increased receptor-mediated cellular internalization of the ADC. The data were represented as mean ± SEM (n = 3). ( c ) Comparison of the cell viability of LT-025, free antibody, and the KIM-1 MMAE ADC by MTT assay. The data showed significantly higher cell death by LT-025 in comparison to free KIM-1 antibody and KIM-1 MMAE ADC. The data were represented as mean ± SEM (n = 3, two-way ANOVA followed by Tukey’s multiple comparison test). ( d ) Confocal images of Renca with and without treatment of LT-025 (100 μg/ml) displaying tubulin disruption. The images showed disruption of the tubulin (green) crosslinking by DM1, a known microtubule inhibitor. Sacale bar = 10 μm. ( e ) Quantification of tubulin in untreated and LT-025 treated cells from the confocal images. The mean pixel intensity of the tubulin (green) was plotted after normalizing with the mean pixel intensity of actin (red). The data were represented as mean ± SEM (n = 23, Unpaired t -test with Welch’s correction). ( f ) Quantification of the area of the control and treated Renca cells. The data showed a significant reduction in the area of the cells. The data were represented as mean ± SEM (n = 23, Unpaired t -test with Welch’s correction). ( g ) The bar plot showing the % of apoptotic cells measured with flow cytometry using Annexin-V AF645 apoptosis kit. The data represent significantly higher apoptosis in normal Renca by LT-025 compared to the control and sunitinib-treated group. (n = 3, one-way ANOVA followed by Tukey’s multiple comparison test). ( h ) The bar plot showing the % of apoptotic sunitinib-resistant Renca measured with flow cytometry using Annexin V-AF645 apoptosis kit. The data represent significantly higher apoptosis in sunitinib-resistant Renca by treatment with dual therapy (LT-025 + sunitinib) compared to the control and monotherapy. 12.5 μM of sunitinib and 100 nM of ADC were used for the study (n = 3, one-way ANOVA followed by Tukey’s multiple comparison test).

Article Snippet: Renca cells (1×10 6 cells in PBS) were injected subcutaneously into flank region of 8-10 weeks syngeneic male balb/c mice (Charles River Laboratories, Strain Code 028).

Techniques: MTT Assay, Comparison, Disruption, Control, Flow Cytometry

Journal: bioRxiv

Article Title: A strategically designed homogeneous antibody-drug conjugate improves safety and therapeutic index in renal cell carcinoma

doi: 10.64898/2026.01.12.699160

Figure Lengend Snippet: Change in E-cadherin amount in Renca cells after treatment with LT-025. The western blot data (left) and its quantification with respect to the GAPDH intensity (right) show an increase in E-cadherin. This signifies a reduction in epithelial-mesenchymal transition (EMT) and lower metastatic potential.

Article Snippet: Renca cells (1×10 6 cells in PBS) were injected subcutaneously into flank region of 8-10 weeks syngeneic male balb/c mice (Charles River Laboratories, Strain Code 028).

Techniques: Western Blot