Journal: International journal of cancer

Article Title: Antitumor effects of histone deacetylase inhibitor on Ewing's family tumors.

doi: 10.1002/ijc.21069

Figure Lengend Snippet: FIGURE 2 – The activity of histone acetyltransferase suppressed by EWS-Fli1 and enhanced by FK228. (a) WE68 cells were treated with RNA interference for 48 hr to knock down EWS-Fli1. Nuclear extracts were extracted from the cells and subjected to HAT assay and Western blot analysis. NT, siScr and siBPEFI represent no treatment, 21 nt scrambled siRNA and 21 nt siRNA corresponding to the sequence of the breakpoint of EWS-Fli1 type I, respectively. siBPEFI treatment suppressed EWS-Fli1 expression and significantly increased HAT activity, whereas siScr treatment did not alter either EWS-Fli1 expression or HAT activity compared with NT. These experiments were repeated at least 3 times. (b) HAT assay using nuclear extracts from WE68 cells treated with FK228 at doses of 2.31–18.5 nM. HAT activity in the FK228-treated cells was dose-dependently increased to up to 30-fold that in nontreated cells. (c) SK-N-MC cells were cul- tured in the absence or presence of 2.31–18.5 nM FK228 for 24 hr. Total cell lysates were extracted from the cells and subjected to West- ern blot analyses using antibodies to p300, CBP, PCAF and TAFII p250. (d) Nuclear extracts of SK-N-MC cells treated with or without 9.25 nM of FK228 were immunoprecipitated with IgG and anti-ace- tyl-Lysine antibody. The immunoprecipitated proteins were separated in a 4–12% gradient gel and blotted with PCAF antibody. Input repre- sents 10% of input of the nuclear extracts. WB, Western blot.

Article Snippet: The antibody used was mouse monoclonal antibody against PCAF (Santa Cruz Biotechnology) at 1:100 dilution.

Techniques: Activity Assay, Knockdown, HAT Assay, Western Blot, Sequencing, Expressing, Immunoprecipitation

Journal: Cancer Medicine

Article Title: Impact of bromodomain‐containing protein 4 (BRD4) and intestine‐specific homeobox (ISX) expression on the prognosis of patients with hepatocellular carcinoma' for better clarity

doi: 10.1002/cam4.4094

Figure Lengend Snippet: PCAF, BRD4 determined by Western blot in anti‐ISX immunoprecipitates of tumor tissues from patients with liver cancer

Article Snippet: The primary antibodies used in this study were PCAF (1:1,000 dilution; #3378S; Cell Signaling Technology), b‐actin (1:10,000 dilution; #4967L; Cell Signaling Technology), GFP (1:500 dilution; SC‐9996; Santa Cruz Biotechnology), ISX (1:200 dilution; sc‐398934; Santa Cruz Biotechnology), BRD4 (1:1,000 dilution; #13440S; Cell Signaling Technology).

Techniques: Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: PCAF Accelerates Vascular Senescence via the Hippo Signaling Pathway

doi: 10.1155/2022/1200602

Figure Lengend Snippet: PCAF were involved in the regulation of endothelial senescence. (a) and (b) Western blotting analysis and qualification of p53, p16, pH2A.X, and p21 protein expression and relative mRNA expression of IL-6 and IL-1 β in PA-treated HUVECs transfected with scramble or PCAF siRNA. (c) and (d) Western blotting analysis and qualification of p53, p16, pH2A.X, and p21 protein expression and relative mRNA expression of IL-6 and IL-1 β in PA-treated HUVECs transfected with vector or PCAF plasmid. (e) and (f) Sa β G staining and quantitative analysis in HUVECs transfected with scramble or PCAF siRNA in the presence of PA or not. Scale bars, 20 μ m. (g) and (h) Sa β G staining and quantitative analysis in HUVECs transfected with vector or PCAF plasmid in the presence of PA or not. Scale bars, 20 μ m. Data are expressed as mean ± SEM ( n = 3 for each experiment. ( ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, n.s. P > 0.05).

Article Snippet: Primary antibody against PCAF (3305, CST), p53 (sc-126, santa cruz), p21 (sc-6246, santa cruz), p16 (sc-166760, santa cruz), Ubiquitin (sc-53509, santa cruz), phosphor-Histone H2A.X(Ser139) (#2577, CST), YAP (#14074, CST), MDM2 (BS-1223, Biogot), phospho-YAP (#13008, CST), TAZ (#83669, CST), phospho-TAZ (#59971, CST), clathrin (ab21679, abcam), caveolae (ab2910, abcam), mTOR (ab2732, abcam), p-mTOR (Ser2448) (ab109268, abcam), p-mTOR(Ser2481) (ab137133, abcam), IL-1 β (sc-12742, snata cruz), IL-6 (sc-130326, santa cruz), GAPDH (#AP0063, Biogot), histone H3(BS3718, Biogot), and PA (P0500, Sigma-Aldrich) were purchased commercially.

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: PCAF Accelerates Vascular Senescence via the Hippo Signaling Pathway

doi: 10.1155/2022/1200602

Figure Lengend Snippet: PCAF mediated PA-induced endothelial senescence through Hippo signaling pathway. (a) KEGG analysis showed that PCAF may regulate endothelial senescence via Hippo-YAP pathway. (b) Western blotting analysis and qualification of phosphorylated YAP expression in HUVECs treated with PA for the indicated time points. (c) Immunofluorescence of YAP localization in HUVECs treated with or without PA. Scar bars, 10 μ m. (d) Expression of total YAP proteins was analyzed by immunoblot in nuclear and cytoplasmic protein extractions from in HUVECs treated with or without PA. (e) and (f): HUVECs were transfected with YAP siRNA or overexpression plasmid. The expression of p53, p16, pH2A.X and p21 protein were by immunoblotting. (g) and (h) HUVECs were transfected with YAP siRNA or overexpression plasmid, IL-6 and IL-1 β mRNA expression were analyzed by RT-PCR. (i) IHC staining and its qualification of vessels from 4w and one and half-year-old mice with YAP antibody. Scale bars, 100 μ m. (j) and (k) HUVECs were transfected with PCAF siRNA or overexpression plasmid, the expression of p-YAP and t-YAP protein were by immunoblotting. (l) HUVECs were transfected with PCAF siRNA or overexpression plasmid treated with or without PA. Expression of YAP proteins was analyzed by immunoblot in nuclear and cytoplasmic protein extractions. (m) Immunofluorescence of YAP localization in HUVECs infected with si PCAF or PCAF plasmid or vector. Scar bars, 10 μ m. (n) and (o) Flag-tagged YAP or Flag-tagged PCAF was transfected into HUVECs before stimulated with PBS or PA for 48 h, Co-IP were performed by using corresponding magnetic beads. t-YAP: total YAP; p-YAP: phosphorylated YAP. Data are expressed as mean ± SEM ( n = 3 for each experiment. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, n.s. P > 0.05).

Article Snippet: Primary antibody against PCAF (3305, CST), p53 (sc-126, santa cruz), p21 (sc-6246, santa cruz), p16 (sc-166760, santa cruz), Ubiquitin (sc-53509, santa cruz), phosphor-Histone H2A.X(Ser139) (#2577, CST), YAP (#14074, CST), MDM2 (BS-1223, Biogot), phospho-YAP (#13008, CST), TAZ (#83669, CST), phospho-TAZ (#59971, CST), clathrin (ab21679, abcam), caveolae (ab2910, abcam), mTOR (ab2732, abcam), p-mTOR (Ser2448) (ab109268, abcam), p-mTOR(Ser2481) (ab137133, abcam), IL-1 β (sc-12742, snata cruz), IL-6 (sc-130326, santa cruz), GAPDH (#AP0063, Biogot), histone H3(BS3718, Biogot), and PA (P0500, Sigma-Aldrich) were purchased commercially.

Techniques: Western Blot, Expressing, Immunofluorescence, Transfection, Over Expression, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry, Infection, Co-Immunoprecipitation Assay, Magnetic Beads

Journal: Oxidative Medicine and Cellular Longevity

Article Title: PCAF Accelerates Vascular Senescence via the Hippo Signaling Pathway

doi: 10.1155/2022/1200602

Figure Lengend Snippet: YAP is the downstream of PCAF in PA-stimulated senescence in HUVECs. (a)–(c) PCAF and YAP were knocked down in HUVECs using siRNA in the treatment of PA or not. The expression of p53, p16, pH2A.X, and p21 protein were by immunoblotting (a), relative IL-6 and IL-1 β mRNA expression were analyzed by RT-PCR (b) and immunoblotting (c). (d) Quantitative analysis of Sa β G staining. Scale bars, 20 μ m. E-G: PCAF and YAP were overexpressed in HUVECs using plasmids in the treatment of PA or not. The expression of p53, p16, pH2A.X, and p21 protein were by immunoblotting (e), relative IL-6 and IL-1 β mRNA expression were analyzed by RT-PCR (f) and immunoblotting (g). (h) Quantitative analysis of Sa β G staining. Scale bars, 20 μ m. Data are expressed as mean ± SEM ( n = 3 for each experiment. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, n.s. P > 0.05).

Article Snippet: Primary antibody against PCAF (3305, CST), p53 (sc-126, santa cruz), p21 (sc-6246, santa cruz), p16 (sc-166760, santa cruz), Ubiquitin (sc-53509, santa cruz), phosphor-Histone H2A.X(Ser139) (#2577, CST), YAP (#14074, CST), MDM2 (BS-1223, Biogot), phospho-YAP (#13008, CST), TAZ (#83669, CST), phospho-TAZ (#59971, CST), clathrin (ab21679, abcam), caveolae (ab2910, abcam), mTOR (ab2732, abcam), p-mTOR (Ser2448) (ab109268, abcam), p-mTOR(Ser2481) (ab137133, abcam), IL-1 β (sc-12742, snata cruz), IL-6 (sc-130326, santa cruz), GAPDH (#AP0063, Biogot), histone H3(BS3718, Biogot), and PA (P0500, Sigma-Aldrich) were purchased commercially.

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: PCAF Accelerates Vascular Senescence via the Hippo Signaling Pathway

doi: 10.1155/2022/1200602

Figure Lengend Snippet: Knockdown of PCAF ameliorated vascular senescence. (a) The concentration of IL-1 β in the plasma from ctrl ( n = 6), aged+AAV9-Luc ( n = 8), and aged+AAV9-PCAF ( n = 7) mice were determined by ELISA kit according to its instruction. (b) and (c) Representative Sa β G staining of aortic sinus sections (b) and its qualification analysis and thoracoabdominal aorta (i). Scale bars, 20 μ m. (d) and (e): The expression of senescence-associated markers (d) and inflammatory factors (e) in the aorta from each group were assessed by immunoblotting. (f) The expression of YAP and p-YAP were investigated by immunoblotting. (g) and (h) The expression of p16, p21 (g) and p-YAP (h) were detected by IHC. Scale bars, 20 μ m. Data are expressed as mean ± SEM ( ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, n.s. P > 0.05).

Article Snippet: Primary antibody against PCAF (3305, CST), p53 (sc-126, santa cruz), p21 (sc-6246, santa cruz), p16 (sc-166760, santa cruz), Ubiquitin (sc-53509, santa cruz), phosphor-Histone H2A.X(Ser139) (#2577, CST), YAP (#14074, CST), MDM2 (BS-1223, Biogot), phospho-YAP (#13008, CST), TAZ (#83669, CST), phospho-TAZ (#59971, CST), clathrin (ab21679, abcam), caveolae (ab2910, abcam), mTOR (ab2732, abcam), p-mTOR (Ser2448) (ab109268, abcam), p-mTOR(Ser2481) (ab137133, abcam), IL-1 β (sc-12742, snata cruz), IL-6 (sc-130326, santa cruz), GAPDH (#AP0063, Biogot), histone H3(BS3718, Biogot), and PA (P0500, Sigma-Aldrich) were purchased commercially.

Techniques: Knockdown, Concentration Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot

Journal: Cancer Cell International

Article Title: Construction and validation of acetylation-related gene signatures for immune landscape analysis and prognostication risk prediction in luminal breast cancer

doi: 10.1186/s12935-025-03920-w

Figure Lengend Snippet: Knock down of KAT2B inhibited the proliferation and invasion of luminal breast cancer cells. ( A ) Survival analysis of KAT2B in luminal breast cancer in TCGA database. HR = 1.82, 95%CI 1.28–2.6, p < 0.0001. ( B ) ROC curve of KAT2B in luminal breast cancer. ( C ) Survival analysis of KAT2B in luminal breast cancer in the SCAN-B database. ( D ) Cell viability assay and proliferation curve showed knock down of KAT2B could inhibit the proliferation of MCF-7 cell. ( E ) Colony formation assay showed knock down of KAT2B could decrease the colony formation ability of MCF-7 cell. The statistic graph was shown in the right. ( F ) Cell viability assay and proliferation curve showed knock down of KAT2B could inhibit the proliferation of T47D cell. ( G ) Colony formation assay showed knock down of KAT2B could decrease the colony formation ability of T47D cell. The statistic graph was shown at right. H-I. Trans-well assay exhibited that knock down of KAT2B could inhibit the invasion ability of MCF7 (H) and T47D (I) cells. The statistics graph was shown at right. *** p < 0.001

Article Snippet: Primary antibodies against KAT2B (PTM Bio, PTM-5711, dilution 1:1000), TAF1L (Proteintech, 55170-1-AP-50UL, dilution 1:1000), and HRP-conjugated glyceraldehyde 3-phosphate dehydrogenase antibody (Proteintech, HRP-60004, dilution 1:5000) were used.

Techniques: Knockdown, Viability Assay, Colony Assay

Journal: Cancer Cell International

Article Title: Construction and validation of acetylation-related gene signatures for immune landscape analysis and prognostication risk prediction in luminal breast cancer

doi: 10.1186/s12935-025-03920-w

Figure Lengend Snippet: Knock down of KAT2B and TAF1L increased lymphocyte infiltration in vitro. A-B. RT-qPCR showed the expression of CD8 + T cell related chemokines in MCF-7 ( A ) and T47D ( B ) cells after silencing of KAT2B. C-D. RT-qPCR showed the expression of CD8 + T cell related chemokines in MCF-7 ( C ) and T47D ( D ) cells after silencing of TAF1L. ( E ) The schematic of the lymphocyte chemotaxis experiment. F-G. The relative lymphocyte chemotaxis rate after knocking down of KAT2B ( F ) and TAF1L ( G ) in MCF-7 and T47D cells. *** p < 0.001

Article Snippet: Primary antibodies against KAT2B (PTM Bio, PTM-5711, dilution 1:1000), TAF1L (Proteintech, 55170-1-AP-50UL, dilution 1:1000), and HRP-conjugated glyceraldehyde 3-phosphate dehydrogenase antibody (Proteintech, HRP-60004, dilution 1:5000) were used.

Techniques: Knockdown, In Vitro, Quantitative RT-PCR, Expressing, Chemotaxis Assay

Journal: Cancer Cell International

Article Title: Construction and validation of acetylation-related gene signatures for immune landscape analysis and prognostication risk prediction in luminal breast cancer

doi: 10.1186/s12935-025-03920-w

Figure Lengend Snippet: Knock down of KAT2B and TAF1L inhibited luminal breast cancer proliferation in vivo. MCF-7, MCF-7-shKAT2B, and MCF-7-shTAF1L were inoculated into the mammary fat pads of Bal/bc nude mice. The tumor picture ( A ), tumor growth curve ( B ), and tumor wight ( C ) showed the effect of sh-KAT2B/TAF1L in vivo. The mouse wight figure ( D ) showed that shKAT2B/TAF1L had no significant effect on the overall state of the mice

Article Snippet: Primary antibodies against KAT2B (PTM Bio, PTM-5711, dilution 1:1000), TAF1L (Proteintech, 55170-1-AP-50UL, dilution 1:1000), and HRP-conjugated glyceraldehyde 3-phosphate dehydrogenase antibody (Proteintech, HRP-60004, dilution 1:5000) were used.

Techniques: Knockdown, In Vivo

Journal: Current Protein & Peptide Science

Article Title: Mutations of Chromatin Structure Regulating Genes in Human Malignancies

doi: 10.2174/1389203717666160122120008

Figure Lengend Snippet: Mutations in histone modification components reported in TCGA publications.

Article Snippet: , KAT2B (PCAF) , Lung , Small Cell Lung Cancer (Johns Hopkins, Nature Genetics 2012) , 2 , 42 , 4.8%.

Techniques: Modification

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, as a lipid metabolism-related biomarker, inhibited RCC progression (A) Triglyceride content statistics in RCC cells. (B) Venn diagram of four datasets. Lipid biosynthetic related genes and lipid metabolic related genes were from GSEA database. (C) Expression levels of IGF1R, AKT1, and KAT2B in clear cell renal cell carcinoma (ccRCC) using TCGA database. (D) The KAT2B protein expression of 72 paired ccRCC and adjacent normal tissues in TCGA-KIRC project. (E-F) Representative images of IHC staining of KAT2B in RCC (E) and statistical analysis (n = 80) (F). (G) Kaplan-Meier survival curves showing the impact of KAT2B expression on overall survival in RCC. (H-I) The tSNE map of RCC tissues ( GSE159115 ) and expression of KAT2B in RCC and normal epithelium cells. (J-K) The mRNA (I) and protein (J) expression levels of KAT2B in HK-2 and RCC cells. (L-M) The cell growth curves of KAT2B overexpression and knockdown RCC cells were determined by CCK8 assays (n = 4). (N-O) Colony formation assays of KAT2B overexpression and knockdown RCC cells. (P) Xenografts from Caki-1 cells with KAT2B overexpression. (Q) The growth curves of RCC with KAT2B overexpression. (R-S) The liver photo and H&E staining of liver tissue in the metastatic model. Data were analyzed by unpaired t test (A, L, Q), paired t test (D, F), wilcoxon test (C), or one-way ANOVA (J,M).

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry, Over Expression, Knockdown, 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: KAT2B suppressed de novo lipogenesis (A) Gene set enrichment analysis (GSEA) assays for the correlations between the KAT2B and fat cell differentiation and lipid metabolism according to the TCGA-KIRC project. FDR < 25 % and P < 0.05 were considered statistically significant. (B) Heatmap of down-regulated lipid in Caki-1 cells with KAT2B overexpression (fold_chang > 1.35, variableimportance inprojection (VIP) > 1 and P_value < 0.05). (C-D) Representative oil red O staining images of RCC cells with KAT2B wild or enzymatically dead form stable overexpression or knockdown, and statistical analysis (n = 3, independent‐samples t‐test). (E-F) Relative TG content of RCC cells with KAT2B stable overexpression or knockdown (n = 4). (G) Representative oil red O staining images of xenografts with KAT2B overexpression. (H) Relative TG content of xenografts with KAT2B overexpression (n = 5). (I) Schematic diagram of construction of spontaneous kidney tumors with KAT2B knockdown. (J) H&E and oil red O staining of spontaneous kidney tumors with KAT2B knockdown, and statistical analysis (n = 5). (K) KAT2B protein levels in spontaneous kidney tumors with KAT2B knockdown were identified. (L) Mechanism diagram of lipid droplet homeostasis. (M) After KAT2B knockdown, the cells were cultured in a medium with 10 % lipid free fetal bovine serum to eliminate the impact of lipid absorption. 5uM chloroquine and 1uM atglistatin were employed to inhibit lysosomal function and ATGL activity, respectively, thereby suppressing lipolysis. After 48 h, the cells were fixed and lipid droplets were visualized using bodipy staining for statistical analysis (n = 5, independent‐samples t‐test). Data were analyzed by unpaired t test (E, H, J), or one-way ANOVA(C, D, F, M).

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Cell Differentiation, Over Expression, Staining, Knockdown, Cell Culture, Activity Assay

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: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

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: KAT2B acetylated HDAC5 at K726 B) Venn diagram identifying proteins that interact with KAT2B and potentially regulate lipid metabolism (A). Among the four candidates, HDAC5 has the highest binding affinity with KAT2B (B). (C) Endogenous Co-Immunoprecipitation (Co-IP) experiments identified the interaction between KAT2B and HDAC5 in RCC cells. (D) The exogenous interaction between KAT2B and HDAC5 was determined by Co-IP assays using Flag and Myc antibodies in 293 T cells. (E) Schematic representation of full-length and truncated mutants of the KAT2B-FLAG structure. (F) FLAG-tagged full-length or truncated mutants of KAT2B were expressed in 293 T cells. Extracts were immunoprecipitated with anti-Flag or anti-HDAC5 antibodies, and bound HDAC5 or Flag was examined by western blots using anti-HDAC5 or anti-Flag antibodies. (G) Western blots were used to assess HDAC5 expression and its acetylation levels in RCC cells with KAT2B overexpression. (H) Mass spectra of the K726 site acetylation of HDAC5 after overexpression of KAT2B in 293 T cells. (I) Comparison of K726 and its surrounding residues between different species. (J) Following the overexpression of KAT2B and either wild-type HDAC5 or a K726 site mutant plasmid, the acetylation levels of HDAC5 were assessed. (K) Purified wild-type and acetyltransferase-inactive mutant KAT2B proteins were co-incubated with purified wild-type HDAC5 and K726R mutant proteins in a buffer containing acetyl-CoA. The acetylation level of HDAC5 was analyzed by western blot. (L) Representative fluorescence image of HDAC5 in RCC cells with KAT2B (wild or dead) overexpression and statistical diagram of nuclear and cytoplasmic distribution. (M) The ratio of HDAC5 nuclear fluorescence intensity to cytoplasmic fluorescence intensity in RCC with KAT2B or KAT2B dead overexpression (n = 10). (N) Nuclear HDAC5 expression in RCC cells with KAT2B (wild or dead) overexpression was assessed using western blot. Data were analyzed byone-way ANOVA (M).

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Expressing, Over Expression, Comparison, Mutagenesis, Plasmid Preparation, Purification, Incubation, Fluorescence

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 destroyed HDAC5/LSD1 complex assembly and suppressed FASN transcriptional activity Co-IP assays were performed to verify the interaction strength between wild-type HDAC5 or the K726R mutant and Exportin1 with KAT2B overexpression. (B) Representative immunofluorescence images of wild-type HDAC5, K726R mutant HDAC5, and NES-deleted HDAC5 with KAT2B overexpression in RCC cells. (C-D) Western blots were used to assess HDAC5 and LSD1 expression in RCC cells with KAT2B overexpression or knockdown. (E) The interaction between HDAC5 and LSD1 was determined by Co-IP assays in RCC cells. (F) The interactions between HDAC5 (wild, K726Q, and K726R) and LSD1 were determined by Co-IP assays in 293 T cells. (G) RCC cells were treated with Eltanexor (60 nM) to inhibit Exportin1 activity. The levels of nuclear HDAC5, total HDAC5, Exportin1, and LSD1 were detected using Western blot. (H) Protein stability experiment of LSD1 in RCC cells with KAT2B overexpression after treated with 100 μM cycloheximide (CHX) for 0 h, 1 h, 2 h, 3 h, and 4 h and statistical diagram. (I) Following the addition of chloroquine (10 μM) or MG132 (8 μM) to RCC, LSD1 protein expression was assessed. (J) RCC cells with KAT2B overexpression were immunoprecipitated with LSD1 antibody, and the level of ubiquitin was detected. (K) LSD1 and FASN expression were detected in RCC cells with KAT2B (wild or dead) and/or HDAC5 (wild, 726Q or 726R) overexpression. (L) Schematic diagram illustrating KAT2B-mediated acetylation of HDAC5, promoting its cytoplasmic mislocalization, which resulted in the disruption of the HDAC5-LSD1 complex in the nucleus and subsequent LSD1 degradation.

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Activity Assay, Co-Immunoprecipitation Assay, Mutagenesis, Over Expression, Immunofluorescence, Western Blot, Expressing, Knockdown, Immunoprecipitation, Ubiquitin Proteomics, Disruption

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: The KAT2B/HDAC5/LSD1/FASN axis repressed RCC lipogenesis and progression in vivo B) The picture of xenografts using Caki-1 cells with KAT2B and/or HDAC5 stable overexpressing. The tumor weight was used for statistical comparison (n = 5). (C) The tumor volume of each group was measured every six days (n = 5). (D) Representative of immunohistochemical (IHC) staining for KAT2B, HDAC5, LSD1, FASN and Ki67 in tumor xenografts. (E) Oil red O staining of the tumor xenografts with KAT2B and/or HDAC5 overexpression. (F) Living fluorescence images of mice in the metastasis model. (G-H) The liver photo and H&E staining of liver tissue in the metastatic model. Data were analyzed by one-way ANOVA (B,C).

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: In Vivo, Comparison, Immunohistochemical staining, Immunohistochemistry, Staining, Over Expression, Fluorescence

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: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Expressing, Sequencing, Methylation, Knockdown

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: Therapeutic targeting of KAT2B-low RCC with a FASN inhibitor (A-B) Representative images of IHC staining of FASN in RCC cohort and statistical analysis. (C) Representative images of IHC staining for FASN and KAT2B in RCC tissues with high and low KAT2B expression. (D) Scatter plot of the relationship among KAT2B expression and FASN expression in advanced RCC tumors (n = 53). (E) The cell viability of ACHN and Caki-1 cells after treated with TVB-2640 (n = 4). (F) The cell viability of 786O and 769P cells after treated with TVB-2640 (n = 10). Proteins from three independent sites in RCC tissues were extracted to detect KAT2B expression. (H) Representative images of Caki-1 and ACHN organoids after treatment with TVB-2640 (7.5 μM). 15 organoids were randomly selected from each group for statistical analysis. (I) Representative images of Caki-1 and ACHN organoids after treatment with TVB-2640 (7.5 μM) (n = 15). (J) Representative images of two PDOs with different KAT2B expression after treatment with TVB-2640 (n = 10). (K) Representative images of PRO-1 staining of PDOs. (L-M) The cell viability of ACHN cells (L) and case 1 primary RCC cells (M) with KAT2B knockdown after treated with TVB-2640 (n = 4). (N-O) The picture (N) of xenograft using 786O cells with KAT2B knockdown after treated with TVB-2640, and tumor growth curve (n = 4). Data were analyzed by unpaired t test (B, H, I, J), one-way ANOVA (O) or two-way ANOVA (E, F, G, L, M).

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Immunohistochemistry, Expressing, Staining, Knockdown

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: Graphic abstract of this research TET1-mediated promoter hypermethylation in RCC leaded to decreased KAT2B expression. Mechanistically, KAT2B acetylated HDAC5 at the K726 site and promoted its nucleus export, thereby failing to form a complex with LSD1 in nucleus. This leaded to increased histone methylation levels and decreased FASN expression, ultimately inhibiting lipogenesis and RCC progression. FASN inhibition might be useful in treating KAT2B-low RCC progression by targeting de novo lipogenesis.

Article Snippet: KAT2B overexpression and HDAC5 overexpression lentivirus, KAT2B‐targeted shRNA lentivirus, and overexpression plasmids of LSD1 and KAT2B (wild and enzyme-inactivated) was provided by Genechem Co. Ltd (China).

Techniques: Expressing, Methylation, Inhibition