rt112 cells  (ATCC)

Journal: bioRxiv

Article Title: CAR T cells targeting Nectin-4 safely overcome resistance to anti-Nectin-4 antibody-drug conjugate in solid tumors

doi: 10.1101/2025.09.30.679440

Figure Lengend Snippet: A. Nectin-4 expression on three cell lines in comparison with SUM190PT. B. MCF7, HT29, RT112 and SUM190PT cell lines were treated with CD30-MMAE(negative control) and enfortumab vedotin. MCF7, HT29 and RT112 cell lines are resistant to enfortumab vedotin whereas SUM190PT is sensible. n = 2 biologic replicates were examined in one experiment. C . 4-hour caspase 3/7 cytotoxicity assay indicating a tumor killing of CARTN4 cells against MCF7, HT29, RT112. n=4 healthy donors were examined in three independent experiments. Data in panels B, and C are presented as Mean +/- SEM. P-values were calculated using a 2way ANOVA. Source data are provided in the Source Data file.

Article Snippet: RT112 cell line was purchased from DSMZ Cell Dive (Braunschweig, Germany).

Techniques: Expressing, Comparison, Negative Control, Cytotoxicity Assay

Journal: Cell Death Discovery

Article Title: CDK4/6 inhibition initiates cell cycle arrest by nuclear translocation of RB and induces a multistep molecular response

doi: 10.1038/s41420-024-02218-6

Figure Lengend Snippet: T24 cells were treated with 1µM PD and harvested at 8, 16, 20, 24, and 48h for RNA sequencing. A PCA shows replicate clustering and separation of time points. PC Principal component. B Heatmap displaying the Log2 fold change of gene expression at the indicated timepoints, compared to the untreated 0-hour control. Rows are clusterd based on Euclidean distance. C Heatmap displaying the scaled (0–1) normalized enrichment scores of KEGG pathways at the indicated time points compared to the untreated 0-hour control. K-mean clustering revealed four major clusters of differentially expressed pathways. D – G CDK4/6 inhibition differentially regulates signaling pathways following four major distinct patterns. KEGG pathways were scanned for pathways that fit one of the four major expression patterns observed in the pathway enrichment analysis. H Time kinetics based on normalized enrichment score compared with the untreated 0-hour control for cell cycle-related pathways of KEGG and GOBP database. I , J RT112 and T24 cells were analyzed by flow cytometry at the indicated time points after treatment with 1µM PD. Cell cycle progression was assessed by EdU incorporation and 7-AAD staining. Values are ± SD as a percentage of total cells and represent three independent replicates. NES: normalized enrichment score.

Article Snippet: RT112 cells were obtained from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and maintained in RPMI medium containing 10% FBS, 1% NEEA and 1% penicillin/streptomycin at 37°C in 5% CO 2 .

Techniques: RNA Sequencing, Gene Expression, Control, Inhibition, Protein-Protein interactions, Expressing, Flow Cytometry, Staining

Journal: Cell Death Discovery

Article Title: CDK4/6 inhibition initiates cell cycle arrest by nuclear translocation of RB and induces a multistep molecular response

doi: 10.1038/s41420-024-02218-6

Figure Lengend Snippet: A T24 and RT112 cells were treated with 1µM PD for 0, 0.5, 4, 8, 12 or 24h. Protein expression was analyzed by immunoblotting. B T24 and RT112 cells were treated with 5µg/ml or 20µg/ml CHX alone or in combination with 1µM PD for 0, 4, 8, 12 or 24h. Protein expression was assessed by immunoblotting. A plus sign indicates presence, while a minus sign indicates absence. C RB signals in Fig. 3 A and B were quantified densitometrically, normalized to the respective housekeeping protein and displayed relative to 0h. Asterisks indicate a significant difference to the respective 0-hour control. D T24 and RT112 cells were treated with 1µM PD for 24h, both with and without MG-132 and epoxomicin as the indicated concentrations. Protein expression was assessed by immunoblotting. E T24 and RT112 cells were transfected with control or gankyrin siRNA oligos. After 24h, cells were treated with 1µM PD for 8 or 24h and Western blot analysis was performed using antibodies against RB, gankyrin and GAPDH. RB signals were quantified densitometrically and expressed as the ration si-gankyrin/si-ctrl at each time point after normalized to the respective housekeeping protein. F RT112 cells transfected with pCMV HA hRB-wt (RB-WT), pCMV HA hRb delta CDK (RB-ΔCDK) or pCMV HA mpRB (RB-mpRB) plasmids for 24h were then treated with 1µM PD for the indicated hours. Exogenous RB protein was detected by Western blotting. RB signals were quantified densitometrically and expressed as the ratio relative to 0h after normalized to the respective housekeeping protein. Error bars indicate SD, asterisks indicate ANOVA. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: RT112 cells were obtained from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and maintained in RPMI medium containing 10% FBS, 1% NEEA and 1% penicillin/streptomycin at 37°C in 5% CO 2 .

Techniques: Expressing, Western Blot, Control, Transfection

Journal: Cell Death Discovery

Article Title: CDK4/6 inhibition initiates cell cycle arrest by nuclear translocation of RB and induces a multistep molecular response

doi: 10.1038/s41420-024-02218-6

Figure Lengend Snippet: A Differential gene expression analysis of Cul1, SKP1 and RBX1 at indicated time points compared to 0-hour control. B Immunoblotting of RB was performed in RT112 and T24 cells treated with 1µM PD for 8h, either in combination with or without 0.2μM MLN4924. RB protein was quantified densitometrically, normalized to GAPDH and the value after PD treatment was compared to the control. C T24 and RT112 cells were treated with 1µM PD either in combination with or without 0.2µM MLN4924 for 8h. Cell cycle analysis was assessed by EdU incorporation and 7-AAD staining. D T24 and RT112 cells were transfected with control, Cullin 1–4 or Cullin 1–5 siRNA oligos. After 24h, the cells were treated with 1µM PD for 8h, and Western blot analysis was conducted using antibodies against RB, pRB and Cul1. After densitometric analysis, normalized to GAPDH, the values of siRNA were referred to the control. E , F Dose–response curves were generated for PD or MLN4924 monotherapy in RT112 and T24 cells. G , I A dose–response matrix was constructed for combination therapy. H , J An interaction landscape was created based on the dose-response matrix. The ZIP synergy score was calculated for the combination therapy. The dose–response data represent the mean from at least 3 biological replicates. Error bars indicate SD, asterisks indicate ANOVA.* P < 0.05; ** P < 0.01; *** P < 0.001, **** P < 0.0001, ns, not significant.

Article Snippet: RT112 cells were obtained from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and maintained in RPMI medium containing 10% FBS, 1% NEEA and 1% penicillin/streptomycin at 37°C in 5% CO 2 .

Techniques: Gene Expression, Control, Western Blot, Cell Cycle Assay, Staining, Transfection, Generated, Construct

Journal: Cell Death Discovery

Article Title: CDK4/6 inhibition initiates cell cycle arrest by nuclear translocation of RB and induces a multistep molecular response

doi: 10.1038/s41420-024-02218-6

Figure Lengend Snippet: A Differential gene expression analysis was performed for the RNAseq data. The fold change in expression for KPNAs and KPNB1 after 8h compared to the 0-hour control was displayed. B T24 cells were treated with 1µM PD for 8h. RT-qPCR was performed to analyze the mRNA expression of these genes. C Subcellular localization of RB and pRB proteins in T24 and RT112 cells harvested at indicated time points after 1µM PD treatment was analyzed by Western blot; RB levels are shown as a ratio to the untreated control at 0h after densitometric quantification of Western blot signals. D T24 cells were treated with 1µM PD for 0, 8 or 24h. Subcellular localization of RB and pRB was assessed by immunofluorescence staining. Scale bar equals 20µm. E RT112 cells transfected with RB-WT or deleted NLS motif (RB-dNLS) plasmids and were treated with 1µM PD for the indicated time points. Expression of endogenous and exogenous RB was determined by Western blotting. Exogenous RB was quantified by densitometry and displayed relative to the untreated 0-hour control. F RT112 cells were transfected with RB-WT or RB-dNLS plasmids and then treated with 1µM PD for 8h. Subcellular localization of RB and pRB proteins were analyzed by Western blotting. Exogenous RB was quantified by densitometry and displayed relative to the untreated 0-hour control. G RT112 cells were transfected with RB-WT or RB-dNLS plasmids and then treated with 1µM PD for 8h. Cell cycle analysis was assessed by BrdU incorporation and flow cytometry. The analysis of BrdU-positive cells was performed for the ratio after PD treatment compared to the corresponding untreated cells. All quantifications were performed using data (mean±SD) from three independent experiments. Asterisks indicate ANOVA: * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: RT112 cells were obtained from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and maintained in RPMI medium containing 10% FBS, 1% NEEA and 1% penicillin/streptomycin at 37°C in 5% CO 2 .

Techniques: Gene Expression, Expressing, Control, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Transfection, Cell Cycle Assay, BrdU Incorporation Assay, Flow Cytometry

Journal: Microbiome

Article Title: Dietary fibre supplementation enhances radiotherapy tumour control and alleviates intestinal radiation toxicity.

doi: 10.1186/s40168-024-01804-1

Figure Lengend Snippet: Fig. 5 Bacterial supernatants from the cocultures of B. acidifaciens and F. prausnitzii conferred stronger anti-tumour phenotypes in bladder cancer cells. a The five bacterial supernatants used in this experiment. b Western blot analysis of histone acetylation (N = 3) of RT112 cells treated with different bacterial supernatants. Histone acetylation levels were determined after treating with 100 mL bacterial supernatants in 2 mL of medium for 24 h. c The cell survival of RT112 cells treated with 200 mL of GAM broth or bacterial supernatants in 500 mL of medium for 2 days (N = 3). d Immunofluorescence microscopy analysis of γ-H2AX levels (N = 3) in RT112 cells treated with 100 mL bacterial supernatants in 2 mL of medium for 24 h. DNA damage was evaluated after treating with 2 Gy IR. Kruskal-Wallis test and Dunn’s multiple comparison tests were conducted to compare the means of different groups. e Linear quadratic survival curves of RT112 cells treated with 100 or 400 μL bacterial supernatant from BA+FP for 24 h before receiving irradiation of 0–8 Gy (N = 3). b, c and e One-way ANOVA with Bonferroni’s multiple comparison test was used to compare the means of different dietary groups. f Principal component analysis of known metabolites of different bacterial supernatants. The clustering effect was assessed using the ADONIS test (R2 = 0.6495, Pr(>F) = 0.001). g Relative levels of metabolites enriched in the bacterial supernatant of BA+FP. ANOVA test, followed by post-hoc analysis using Fisher’s LSD and p value adjustment using the Benjamin-Hochberg method, was applied to identify the metabolites with significantly different levels across the groups. pHs of GAM broth and bacterial supernatants were all neutralised to 7.2. BA+FP denotes the co-culture of B. acidifaciens and F. prausnitzii, while Bif+FP denotes the co-culture of Bifidobacterium and F. prausnitzii. Data are presented as mean ± SD

Article Snippet: The RT112 human bladder carcinoma cell line was obtained from DSMZ (Germany) and cultured in RPMI1640 medium (Sigma) supplemented with 10% fetal bovine serum (Invitrogen).

Techniques: Western Blot, Immunofluorescence, Microscopy, Comparison, Irradiation, Co-Culture Assay