a498  (ATCC)

Journal: Journal of Advanced Research

Article Title: Epigenetically silenced KAT2B suppresses de novo lipogenesis through destroying HDAC5/LSD1 complex assembly in renal cell carcinoma

doi: 10.1016/j.jare.2025.08.007

Figure Lengend Snippet: KAT2B suppressed lipogenesis through FASN (A) Key rate-limiting enzymes in de novo lipogenesis and their expression levels in ccRCC and pRCC using TCGA-KIRC and TCGA-KIRP databases. Red squares and blue squares represented genes whose expression were up-regulated or down-regulated in tumors. (B) Schematic diagram for screening key lipid synthesis factors downstream of KAT2B. (C) Statistical analysis of oil red O stainging in 786O cells following knockdown of 10 key lipogenesis factors (n = 3). (D-E) Representative IHC staining for FASN in RCC cohort and statistical analysis (n = 80, paired t‐test). (F-G) After KAT2B knockdown in 786O and ACHN cells, the mRNA and protein expression of FASN was observed. (H) The cell growth curves of A498 and Caki-1 cells with KAT2B and/or FASN overexpression were determined by CCK8 assays (n = 4, independent‐samples t‐test). (I) The relative TG levels in A498 and Caki-1 cells with KAT2B and/or FASN overexpression (n = 4, independent‐samples t‐test). (J) Representative images of oil red O staining of A498 and Caki-1 cells with KAT2B and/or FASN overexpression and statistical analysis (n = 3, independent‐samples t‐test). Data were analyzed by unpaired t test (G), paired t test (E), one-way ANOVA (H, I, J) or two-way ANOVA (C).

Article Snippet: The HK‐2, 293 T, A549, PC9, T47D, MCF7, A498, Caki-1, OSRC-2, 786O, 769P and ACHN cell lines were obtained from the American Type Culture Collection (ATCC, USA) and were cultivated under proper conditions according to the manufacturer’s protocols.

Techniques: Expressing, Knockdown, Immunohistochemistry, Over Expression, Staining

Journal: Journal of Advanced Research

Article Title: Epigenetically silenced KAT2B suppresses de novo lipogenesis through destroying HDAC5/LSD1 complex assembly in renal cell carcinoma

doi: 10.1016/j.jare.2025.08.007

Figure Lengend Snippet: Hypermethylation but not VHL/HIF axis resulted in low expression of KAT2B in RCC Expression levels of HIF2a and KAT2B after hypoxia in RCC cells. (B) Expression level of KAT2B after overexpressing HIF2a in Caki-1 cells. (C) Expression level of KAT2B after overexpressing VHL in A498 cells. (D) RNA stability experiment of KAT2B in RCC and HK2 cells after treated with 20 μg/ml cycloheximide (CHX) for 0 h, 1 h, 2 h, 3 h, and 4 h and statistical diagram. (E-F) Prediction analysis of CpG islands in the sequence range of 3500 bp upstream from the transcriptional start site in the KAT2B promoter region ( http://www.urogene.org/ ). (G-H) The promoter methylation level of KAT2B in ccRCC using online database UCSC Xena ( http://xena.ucsc.edu/ ) and UALCAN ( http://ualcan.path.uab.edu/ ). (I) Scatter plot of the relationship among KAT2B expression and its promoter methylation level. (J) Representative MSP results of KAT2B methylation status in 5 paired adjacent tissues (N) and RCC tissues (T). (K-L) The mRNA and protein levels of KAT2B in RCC cell lines after 5-AZA treatment (n = 3). (M) Scatter plot of the relationship among KAT2B expression and TET1, TET2, and TET3 expression. (N) The KAT2B mRNA expression after knockdown of TET1, TET2, or TET3 in 786O cells. (O) The KAT2B protein expression after TET1 knockdown in RCC cells. Data were analyzed by one-way ANOVA (K,N) or two-way ANOVA (D).

Article Snippet: The HK‐2, 293 T, A549, PC9, T47D, MCF7, A498, Caki-1, OSRC-2, 786O, 769P and ACHN cell lines were obtained from the American Type Culture Collection (ATCC, USA) and were cultivated under proper conditions according to the manufacturer’s protocols.

Techniques: Expressing, Sequencing, Methylation, Knockdown

Journal: Science Advances

Article Title: Genetically engineered human cell–based microrobots for selective cancer cell death

doi: 10.1126/sciadv.aea9831

Figure Lengend Snippet: ( A ) Schematic illustration of the experimental setup with four groups: (i) unmodified 293T cells, (ii) 293T cells modified with GFP only, (iii) 293T cells modified with TRAIL + GFP , and (iv) cell-based microrobots—293T cells modified with TRAIL + GFP and then conjugated to magnetic Janus particles. 293T cells were seeded in the upper chamber of a transwell system; healthy hAMSCs or A498 cancer cells were seeded in the lower chamber. Cell viability was assessed after 3 days of coculture. Created in BioRender. N. O. Dogan (2026), https://biorender.com/3lke7fg . ( B ) TRAIL secretion was measured in all groups at 48 hours posttransfection. Data are presented as the means ± SD from n = 3 technical replicates. One-way ANOVA with Tukey’s post hoc test; n.s. indicates P ≥ 0.05, and **** P < 0.0001. ( C ) CellTiter-Glo viability assays showing a minimal effect on healthy hAMSC viability across groups, while A498 cancer cells exhibited significant death in both TRAIL + GFP and TRAIL + GFP + particles groups. Data are presented as the means ± SD from n = 10 technical replicates. One-way ANOVA with Tukey’s post hoc test; n.s. indicates P ≥ 0.05; * P < 0.05, ** P < 0.01, and **** P < 0.0001. ( D ) Representative live/dead fluorescence images confirming the selective cytotoxicity of cell-based microrobots on A498 cancer cells, with no detectable harm on healthy hAMSCs. Scale bars, 100 μm.

Article Snippet: ACHN and A498 human renal adenocarcinomas (ATCC) and ONCO-DG-1 human ovarian adenocarcinomas (DSMZ) were cultivated in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco).

Techniques: Modification, Fluorescence

Journal: Science Advances

Article Title: Genetically engineered human cell–based microrobots for selective cancer cell death

doi: 10.1126/sciadv.aea9831

Figure Lengend Snippet: ( A ) Differential interference contrast imaging demonstrating the magnetic guidance of cell-based microrobots toward a 3D tumor spheroid. Scale bars, 100 μm. ( B ) As shown by live/dead fluorescence imaging, conditioned medium from cell-based microrobots induced increased tumor cell death in 3D A498 renal adenocarcinoma tumor spheroids, unlike medium from unmodified 293T cells. Scale bars, 100 μm. ( C ) CellTiter-Glo 3D assay revealed significantly reduced tumor spheroid viability after 24 hours of incubation with conditioned medium from cell-based microrobots compared to medium from unmodified 293T cells. Data are presented as the means ± SD, n = 6 technical replicates. One-way ANOVA with Tukey’s post hoc test; **** P < 0.0001.

Article Snippet: ACHN and A498 human renal adenocarcinomas (ATCC) and ONCO-DG-1 human ovarian adenocarcinomas (DSMZ) were cultivated in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco).

Techniques: Imaging, Fluorescence, Incubation