rt112 cell line  (European Collection of Authenticated Cell Cultures)

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: 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

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: YEATS4 is essential for the tumor growth of bladder cancer cells. A) Representative immunohistochemical images of YEATS4 in 78 BC tissues. Scale bar, 100 µm. B) The relative YEATS4 protein level in paired tumor tissues (Tumor) and adjacent normal tissues (Normal) used in (A). C) Overall survival curves were analyzed based on YEATS4 protein levels as measured in (A and B) in BC patients. D) Western blotting of the indicated proteins in the indicated stable cells of 3 independent experiments. E) Cell viability of T24 and UMUC3 cells expressing YEATS4‐targeting sgRNAs was evaluated by MTT assay at the indicated time points. n = 3 independent experiments. F) Colony formation assays were performed for the indicated stable cells. Colony numbers were quantified using ImageJ software. n = 3 independent experiments. G–J) The indicated stable cells as shown in (D) were subcutaneously injected into nude mice. Tumor weights (G and I) and tumor volumes (H and J) were measured. n = 8 nude mice per group were used for T24 experiments, and n = 6 nude mice per group were used for UMUC3 experiments. Data in E‐J are presented as the mean ± SD. P values in C and E–J were calculated using the two‐tailed Student's t ‐test. p values in C were analyzed using the Kaplan‒Meier plots.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Immunohistochemical staining, Western Blot, Expressing, MTT Assay, Software, Injection, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: Depletion of YEATS4 inhibits DNA repair and induces cellular senescence. A) Schematic illustration of the GFP‐based HR reporter assay (DR‐GFP) and NHEJ reporter assay (EJ5‐GFP). B) Relative DNA repair efficiency in UMUC3 cells stably expressing the indicated sgRNAs. n = 3 independent experiments. C and D) Representative immunofluorescence images and quantification data of Rad51 foci (C) and 53BP1 foci (D) formation in T24 stable cells at 4 h after exposure to 10 Gy irradiation. n = 50 cells were analyzed for each group. Scale bar, 10 µm. E) γ‐H2AX levels in the indicated stable cells were analyzed by Western blotting. n = 3 independent experiments. F) Representative immunofluorescence images and quantification data of γ‐H2AX foci and micronuclei (MN) formation (pointed by arrows) in UMUC3 stable cells. n = 50 cells were analyzed for each group. Scale bar, 10 µm. G) Representative images and quantification data of senescence‐associated β‐galactosidase (SA‐β‐gal) staining of the indicated stable cells. n = 3 independent experiments. Data in B, C, D, F, and G are presented as the mean ± SD. P values were calculated using the two‐tailed Student's t ‐test.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Reporter Assay, Stable Transfection, Expressing, Immunofluorescence, Irradiation, Western Blot, Staining, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: HUWE1 is an E3 ligase for the ubiquitination and degradation of YEATS4. A) Schematic of the YEATS4 Protein Stability Regulators Screening Assay (ProSRSA). B) Top 10 candidate genes of MAGeCK analysis. The x‐axis represents the ranking of genes, and the y‐axis indicates the robust rank aggregation (RRA) score of each gene. C) T24 cells were subjected to co‐IP by using anti‐YEATS4 or anti‐IgG to detect endogenous HUWE1. D) The indicated cells were transfected with HUWE1‐targeted siRNAs for 48 h and then were lysed and analyzed by Western blotting. E) HEK293T cells were transfected with indicated plasmids at the indicated concentration for 24 h and then were lysed and analyzed by Western blotting. F and G) T24 cells transfected with HUWE1‐targeted siRNAs for 48 h were treated with 40 µg mL −1 cycloheximide (CHX) at the indicated time points and then lysed and analyzed by Western blotting (F). Quantitation of YEATS4 protein levels was based on the Western blotting results (G). H) T24 cells transfected with HUWE1‐targeted siRNAs for 48 h were subjected to co‐IP with the indicated antibodies followed by Western blotting. I and J) HEK293T cells cotransfected with the indicated plasmids for 12 h were treated with 40 µg/ml CHX at the indicated time points, and then were lysed and analyzed by Western blotting (I). Quantification of YEATS4 protein levels was based on the Western blotting results (J). K) HEK293T cells cotransfected with the indicated plasmids for 48 h were incubated with 10 µM MG132 for 6 h, and then subjected to IP using Ni‐NTA beads followed by Western blotting. Ni‐NTA, nitrilotriacetic acid. Data are representative of n = 3 independent experiments. Data in G and J are presented as the mean ± SD. P values were analyzed using the two‐tailed Student's t ‐test.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Ubiquitin Proteomics, Screening Assay, Co-Immunoprecipitation Assay, Transfection, Western Blot, Concentration Assay, Quantitation Assay, Incubation, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: KAT8 inhibits ubiquitination‐mediated proteasomal degradation of YEATS4. A) Schematics of the proximity labeling assay using YEATS4‐V5‐TurboID‐construct. B) HEK293T cells were cotransfected with the indicated constructs for 48 h, and then subjected to IP using HA‐agarose beads followed by Western blotting. C) T24 cells were subjected to co‐IP by using anti‐YEATS4 or anti‐IgG to detect endogenous KAT8. D and E) Western blotting of the indicated proteins in the indicated stable cells infected with KAT8‐targeted sgRNAs (D) or treated with or without 50 µM MG149 for 48 h (E). Data are representative of 3 independent experiments. F and G) The indicated stable cells expressing KAT8‐targeted sgRNAs were treated with 40 µg/ml CHX at the indicated time points and then were lysed and analyzed by Western blotting (F). Quantitation of YEATS4 protein levels was based on the Western blotting results (G). H and K) HEK293T cells were cotransfected with the indicated plasmids for 48 h and then subjected to immunoprecipitation using streptavidin beads followed by Western blotting. WCL, whole cell lysate. I and J) HEK293T cells cotransfected with the indicated plasmids for 12 h were treated with 40 µg/ml CHX at the indicated time points and then were lysed and analyzed by Western blotting (I). Quantification of YEATS4 protein levels was based on the Western blotting results (J). L and M) The indicated cells transfected with KAT8‐targeted siRNAs for 48 h (L) or incubated with 50 µM MG149 for 48 h (M) were treated with or without 10 µM MG132 for 6 h, and then cells were lysed and analyzed by Western blotting. Data are representative of n = 3 independent experiments. Data in G and J are presented as the mean ± SD. P values were analyzed using the two‐tailed Student's t ‐test.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Ubiquitin Proteomics, Labeling, Construct, Western Blot, Co-Immunoprecipitation Assay, Infection, Expressing, Quantitation Assay, Immunoprecipitation, Transfection, Incubation, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: Acetylation of YEATS4 by KAT8 is critical for its oncogenic function in bladder cancer cell viability. A) Relative DNA repair efficiency in YEATS4 WT and 3KR cells. B) Western blotting of the indicated proteins in YEATS4 WT and 3KR cells. n = 3 biologically independent experiments. C) T24 and UMUC3 cells were treated with or without 50 µM MG149 for 48 h and then were lysed and analyzed by Western blotting. n = 3 biologically independent experiments. D and E) MTT assays (D) and colony formation assays (E) were performed in YEATS4 WT and 3KR cells. F and G) MTT assay (F) and colony formation assay (G) were performed in T24 and UMUC3 cells treated with DMSO or 50 µM MG149. H and I) Tumor growth of YEATS4 WT and 3KR cells was evaluated in vivo. n = 8 nude mice per group. Tumor weights (H) and tumor volumes (I) were measured. J and K) Mice bearing UMUC3 tumors were randomly divided into the indicated groups ( n = 6 mice per group). DMSO or MG149 (5 mg kg −1 ) was injected intraperitoneally at the indicated time points. Tumor weights (J) and tumor volumes (K) were measured. L) Representative immunohistochemical staining images of both YEATS4 and KAT8 in 78 paraffin‐embedded BC tissues. Scale bar, 100 µm. M) Crosstab shows the distribution of cancer tissues in the 78 bladder cancer tissues used in (L) according to the H‐Score of YEATS4 and KAT8. p < 0.0001, χ 2 tests. R , Spearman correlation coefficient. P values were analyzed using Pearson's chi‐squared test, and the R ‐value was analyzed using Spearman's correlation test. N and O) Overall survival curves were generated based on the protein levels of YEATS4 or KAT8 in the BC tissues used in (L). p value was calculated using the Kaplan‒Meier plots. Data in A and D‐K are presented as the mean ± SD. p values were calculated by two‐tailed Student's t ‐test.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Western Blot, MTT Assay, Colony Assay, In Vivo, Injection, Immunohistochemical staining, Staining, Generated, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting the KAT8/YEATS4 Axis Represses Tumor Growth and Increases Cisplatin Sensitivity in Bladder Cancer

doi: 10.1002/advs.202310146

Figure Lengend Snippet: Disruption of KAT8‐mediated YEATS4 acetylation increases the DDP sensitivity of BC cells. A) Apoptosis in parental WT or 3KR cells treated with 5 µM DDP for 48 h. B) Apoptosis in the indicated cells treated with 2.5 µM DDP, 50 µM MG149, and their combination for 48 h. C) YEATS4 WT cells or 3KR cells were treated with the indicated concentrations of DDP for 48 h, and then cell viability was measured by MTT assay. n = 3 biologically independent experiments. D) The indicated cell lines were treated with the indicated concentrations of DDP, MG149, and their combination for 48 h, and then cell viability was measured by MTT assay. n = 3 biologically independent experiments. E and F) Mice bearing parental WT or 3KR tumors were randomly divided into 2 indicated groups (n = 6 mice per group). PBS or DDP (2 mg kg −1 ) was injected intraperitoneally at the indicated time points. Tumor weights (E) and tumor volumes (F) were measured. G and H) Mice bearing T24 tumors were randomly divided into 4 indicated groups (n = 8 mice per group). DDP (2 mg kg −1 ), MG149 (5 mg kg −1 ), and their combination were injected intraperitoneally at the indicated time points. Tumor weights (G) and tumor volumes (H) were measured. Data in A‐H are presented as the mean ± SD. p values were calculated by two‐tailed Student's t ‐test. The IC50 values in C and D were calculated using GraphPad software with Nonlinear regression, Dose‐response‐Inhibiton, log (inhibitor) versus normalized response Variable slope methods.

Article Snippet: Cell lines T24, UMUC3, 5637, J82, TCCSUP, SCaBER, SW780, HT1376, RT4, and HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC), RT112 were obtained from the European Collection of Authenticated Cell Cultures (ECACC), and the BIU87 cell line was obtained from Kunming Cell Bank, Chinese Academy of Sciences.

Techniques: Disruption, MTT Assay, Injection, Two Tailed Test, Software