Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: YEATS2 and H3K27ac were enriched in the promoter of IL6ST. (A) The YEATS2 relationships with acetylation level of the common lysine sites of H3 histone were detected by Western blot. (B) The binding of YEATS2 with H3K27ac was verified by Co-IP. (C) YEATS2 and 17 DEGs mRNA expression levels were detected by performing RT-qPCR. (D) TSA, a histone deacetylase inhibitor, prominently increased IL6ST expression. (E) YEATS2 regulated the protein expression of IL6ST. (F) H3K27ac enrichment in the promoter of IL6ST was predicted by the ENCODE database. (G) ChIP was used to detect YEATS2 and H3K27ac enrichment in the promoter of CTSL, IL6ST, MAD2L1 and FAM3C. (H) IL6ST knockdown partially reversed the effect of YEATS2 on the migration of TE5. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (I) IL6ST was involved in the regulation of YEATS2 on p105/p50 protein.

Article Snippet: In order to screen out the genes, on which YEATS2 and H3K27ac were both enriched, antibodies of YEATS2 (Proteintech Co.) and H3K27ac (CST Co.) were used to precipitate the DNA fragments.

Techniques: Western Blot, Binding Assay, Co-Immunoprecipitation Assay, Expressing, Quantitative RT-PCR, Histone Deacetylase Assay, Knockdown, Migration

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: TAF15 and KAT5 participated in the regulation of YEATS2 on H3K27ac and IL6ST. (A) Co-IP-based mass spectrum (MS) assays were performed in TE5 cells with YEATS2 overexpression. The second-order spectrum of TAF15 was derived from Co-IP-based MS. (B) GeneMANIA database was used to predict the proteins binding to TAF15. (C) Co-IP was employed to test interactions among YEATS2, TAF15 and KAT5. (D) The subcellular localizations of YEATS2, TAF15 and KAT5 were tested by IF. (E) ChIP was employed to test TAF15 and KAT5 enrichment in the IL6ST promoter region. (F) KAT5 and TAF15 participated in the regulation of YEATS2 on the protein expression level of IL6ST. (G) KAT5 but not TAF15 could increase the level of H3K27ac. (H) KAT5 but not TAF15 could increase H3K27ac enrichment in IL6ST promoter, which was verified by ChIP-qPCR. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: In order to screen out the genes, on which YEATS2 and H3K27ac were both enriched, antibodies of YEATS2 (Proteintech Co.) and H3K27ac (CST Co.) were used to precipitate the DNA fragments.

Techniques: Co-Immunoprecipitation Assay, Over Expression, Derivative Assay, Binding Assay, Expressing, ChIP-qPCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: A proposed working model for YEATS2/TAF15/KAT5 complex regulating NF-κB by H3K27 acetylation activated-IL6ST in ESCC.

Article Snippet: In order to screen out the genes, on which YEATS2 and H3K27ac were both enriched, antibodies of YEATS2 (Proteintech Co.) and H3K27ac (CST Co.) were used to precipitate the DNA fragments.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: The expression and prognostic value of YEATS2 in ESCC. (A) The copy number variations of 508 ESCC tissues were analysed by using GISTIC based on our previous project. (B) The copy number amplifications of YEATS2 in 508 ESCC genomes. (C) Kaplan-Meier was performed to analyze the associations of the copy number of YEATS2 with postoperative survival time for G3-stage ESCC patients. (D) The correlation between expression of YEATS2 mRNA and YEATS2 copy number in 155 ESCC tissues. Statistical significance was assessed by Pearson’s chi-squared test. (E) YEATS2 mRNA expression in ESCC tissues and normal esophageal tissues from RNA-seq data of 155 ESCC patients. (F) YEATS2 protein expression in ESCC tissues and normal esophageal tissues from proteomic sequencing data of 124 ESCC patients. The protein expression values have been normalized and log2-transformed.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: Expressing, RNA Sequencing, Sequencing, Transformation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: Relationship between YEATS2 copy number and clinical characteristics of patients.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: Relationship between YEATS2 protein expression and clinical characteristics of patients.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: YEATS2 stable knockdown inhibited the abilities of ESCC cells to proliferate and migrate in vitro and in vivo . (A) Western blot and RT-qPCR were performed to verify the efficiency of YEATS2 knockdown and overexpression acted by CRISPR/Cas9 and CRISPR/dCas9-SAM system. Data shown are the mean ± SD of three biological replicates. p value was calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (B) MTT was performed to detect YEATS2 effect on the proliferation ability of ESCC cells in vitro , including KYSE150 and TE9. Data shown are the mean ± SD of three biological replicates. p values were calculated by one-way ANOVA or unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (C–D) Colony Formation Assay was performed to detect YEATS2 effect on the colony forming ability of ESCC cells in vitro , including KYSE150 and TE9. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (E–F) Transwell was used to detect YEATS2 effect on the migration ability of ESCC cells in vitro , including KYSE150 and TE9. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (G–H) Scratch Wound Healing was used to detect YEATS2 effect on the wound healing ability of ESCC cells in vitro , including KYSE150 and TE9. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (I) Photographic images and hematoxylin–eosin (HE) staining of tumors from nude mice (n = 3 in each group). (J) The volume change of tumors from tumor-bearing nude mice. Tumor volume was calculated and diameters were measured at a regular interval of 6 days for up to 33 days. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (K) Representative images of lungs from tail-vein lung metastatic models (n = 4 in each group) and HE staining for lungs from tail-vein lung metastatic models.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: Knockdown, In Vitro, In Vivo, Western Blot, Quantitative RT-PCR, Over Expression, CRISPR, Colony Assay, Migration, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: NF-κB signaling pathway mediated YEATS2 function. (A) RNA-seq were performed before and after YEATS2 knockdown on KYSE450 and KYSE180 cells. The volcano maps of DEGs in KYSE180 and KYSE450 from RNA-seq. (B) The KEGG analyses of 221 DEGs derived from RNA-seq. (C) The YEATS2 regulations on the transcriptional activity of NF-κB signaling pathway were verified by Dual-Luciferase Reporter Assay. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (D) The YEATS2 regulations on NF-κB p105/p50 and p-NF-κB p105 levels were verified by Western blot. (E, F, H) LPS weakened the effect of YEATS2 knockdown on the proliferation and migration of KYSE450. Data shown are the mean ± SD of three biological replicates. p values were calculated by one-way ANOVA with * p < 0.05, ** p < 0.01, and *** p < 0.001. (G, H) PDTC weakened the effect of YEATS2 overexpression on the migration of TE5. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: RNA Sequencing, Knockdown, Derivative Assay, Activity Assay, Luciferase, Reporter Assay, Western Blot, Migration, Over Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: YEATS2 and H3K27ac were enriched in the promoter of IL6ST. (A) The YEATS2 relationships with acetylation level of the common lysine sites of H3 histone were detected by Western blot. (B) The binding of YEATS2 with H3K27ac was verified by Co-IP. (C) YEATS2 and 17 DEGs mRNA expression levels were detected by performing RT-qPCR. (D) TSA, a histone deacetylase inhibitor, prominently increased IL6ST expression. (E) YEATS2 regulated the protein expression of IL6ST. (F) H3K27ac enrichment in the promoter of IL6ST was predicted by the ENCODE database. (G) ChIP was used to detect YEATS2 and H3K27ac enrichment in the promoter of CTSL, IL6ST, MAD2L1 and FAM3C. (H) IL6ST knockdown partially reversed the effect of YEATS2 on the migration of TE5. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001. (I) IL6ST was involved in the regulation of YEATS2 on p105/p50 protein.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: Western Blot, Binding Assay, Co-Immunoprecipitation Assay, Expressing, Quantitative RT-PCR, Histone Deacetylase Assay, Knockdown, Migration

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: TAF15 and KAT5 participated in the regulation of YEATS2 on H3K27ac and IL6ST. (A) Co-IP-based mass spectrum (MS) assays were performed in TE5 cells with YEATS2 overexpression. The second-order spectrum of TAF15 was derived from Co-IP-based MS. (B) GeneMANIA database was used to predict the proteins binding to TAF15. (C) Co-IP was employed to test interactions among YEATS2, TAF15 and KAT5. (D) The subcellular localizations of YEATS2, TAF15 and KAT5 were tested by IF. (E) ChIP was employed to test TAF15 and KAT5 enrichment in the IL6ST promoter region. (F) KAT5 and TAF15 participated in the regulation of YEATS2 on the protein expression level of IL6ST. (G) KAT5 but not TAF15 could increase the level of H3K27ac. (H) KAT5 but not TAF15 could increase H3K27ac enrichment in IL6ST promoter, which was verified by ChIP-qPCR. Data shown are the mean ± SD of three biological replicates. p values were calculated by unpaired t tests with * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: Co-Immunoprecipitation Assay, Over Expression, Derivative Assay, Binding Assay, Expressing, ChIP-qPCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: YEATS2 promotes malignant phenotypes of esophageal squamous cell carcinoma via H3K27ac activated-IL6ST

doi: 10.3389/fcell.2025.1497290

Figure Lengend Snippet: A proposed working model for YEATS2/TAF15/KAT5 complex regulating NF-κB by H3K27 acetylation activated-IL6ST in ESCC.

Article Snippet: After being blocked, the PVDF membrane was incubated with primary antibodies overnight at 4°C, including YEATS2 (Proteintech Co., 1:1,000), histone 3 (H3) (CST Co., 1:1,000), H3K9 acetylation (H3K9ac) (CST Co., 1:1,000), H3K14 acetylation (H3K14ac) (PTM Co., 1:1,000), H3K18 acetylation (H3K18ac) (PTM Co., 1:1,000), H3K27ac (CST Co., 1:1,000), NF-κB (PTM Co., 1:1,000), TAF15 (Abcam Co., 1:10,000), IL6ST (Santa Cruz Co., 1:1,000), KAT5 (Proteintech Co., 1:1,000), GAPDH (Proteintech Co., 1:5,000).

Techniques: