Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: Representative immunohistochemical staining of TRIM28, ×400

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Immunohistochemical staining, Staining

Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: Associations between the expression of TRIM28 and clinicopathologic characteristics of the 136 cases of ESCC patients

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Expressing

Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: Associations between the expression of TRIM28, clinicopathological parameters and 5-year overall survival rate of the 136 cases of ESCC patients

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Expressing

Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: Relative expression of TRIM28 protein by Western blot between paired ESCC tissues and NEE tissues (Cells in ESCC can express TRIM28 while there was no expression of TRIM28 in cells of NEE)

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Expressing, Western Blot

Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: The distribution of TRIM28 protein in ESCC and NEE tissues by immunofluorescence stainings, ×400. (TRIM28 protein was stained with green fluorescence(→), which was expressed in the nucleus in ESCC cells and nuclei were counterstained with DAPI (A); Analysis results of the fluorescence intensity (B), P = 0.000) (One-way ANOVA)

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Pathology Oncology Research

Article Title: Expression and Significance of TRIM 28 in Squamous Carcinoma of Esophagus

doi: 10.1007/s12253-018-0558-6

Figure Lengend Snippet: TRIM28 protein expression during cancer progression by IHC analysis

Article Snippet: After washing 5 min in PBS for three times, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-TRIM28 antibody (GTX102226; dilution, 1:200; GeneTex, USA) and followed by incubation with reagent I of HRP-labeled goat anti-rabbit IgG antibody (SP-9001, Zhongshan Goldenbridge Biol\Technology Co., Ltd., Beijing, China) for 30 min at 37 °C.

Techniques: Expressing

Journal: Nucleic Acids Research

Article Title: DICER regulates the expression of major satellite repeat transcripts and meiotic chromosome segregation during spermatogenesis

doi: 10.1093/nar/gkaa460

Figure Lengend Snippet: Antibodies used in the study

Article Snippet: TRIM28 (rabbit) , Bethyl Laboratories, Inc. (A304-145A-M) , IP.

Techniques: Negative Control

Journal: Nucleic Acids Research

Article Title: DICER regulates the expression of major satellite repeat transcripts and meiotic chromosome segregation during spermatogenesis

doi: 10.1093/nar/gkaa460

Figure Lengend Snippet: Epigenetic status of the MSR chromatin is disrupted in Dicer1 knockout testes. ( A ) Chromatin immunoprecipitation (ChIP) with an anti-DICER (DCR) antibody followed by PCR using primers specific for different repeat sequences revealed the association of DICER with the MSR-containing genomic regions. DICER did not associate with minor satellite repeats, various transposon sequences (Line1, SineB1, SineB2, IAP) or ribosomal DNA repeats (rDNA). Rabbit IgG (IgG) was used as a negative control for immunoprecipitation. The primers for Gapdh gene were used as a negative control for PCR. The top right panel shows immunoblotting with an anti-DICER antibody to validate the performance of the antibody in the ChIP assay. ( B ) Immunoprecipitation using antibodies against DICER, TRIM33 and TRIM28 followed by western blotting with the same antibodies validated the interaction of DICER (DCR) with TRIM33 (TR33) and TRIM28 (TR28). LYS, lysate; IP, immunoprecipitation; IgG, control IP with rabbit IgG. ( C ) DICER interacts with histone methyltransferases in the testis. Immunoprecipitation using antibodies against SETDB1 (SET) and SUV39H2 (SUV) followed by immunoblotting with SETDB1, SUV39H2 and DICER antibodies. LYS, lysate; IP, immunoprecipitation; IgG, control IP with rabbit IgG. ( D ) Localization of H3K9me3 appears unaffected in Dicer1 cKO germ cells. PFA-fixed paraffin-embedded controls (CTRL) and Dicer1 cKO testis sections were immunostained with an anti-H3K9me3 antibody. Scale bar: 10 μm. Panels A-C show a representative figure of an experiment that was independently repeated at least two times. ( E ) ChIP with anti-SUV39H2 revealed that the association of SUV39H2 with MSRs is reduced in Dicer1 cKO adult testis. ( F ) Overall H3K9me3 (K9m3) signal on MSR genomic regions appeared reduced in Dicer1 cKO adult testis. The graphs in panels E and F show the quantification of the ChIP-PCR signal (308 bp band) in two independent experiments. The ChIP signal was normalized to the input signal. Signal intensities are shown relative to the CTRL ChIP samples that were set as ‘1’; n = 2, SEM, * P < 0.05. See also and .

Article Snippet: TRIM28 (rabbit) , Bethyl Laboratories, Inc. (A304-145A-M) , IP.

Techniques: Knock-Out, Chromatin Immunoprecipitation, Negative Control, Immunoprecipitation, Western Blot

Journal: bioRxiv

Article Title: A non-canonical EZH2/TRIM28 epigenetic axis drives heparan sulfate remodeling and melanoma metastasis

doi: 10.64898/2026.01.21.700969

Figure Lengend Snippet: (A) Immunoprecipitation of EZH2 in A375 wild-type cells and LC-MS/MS proteomics reveals canonical PRC2 proteins and other known EZH2 interactors (table inset). STRING-DB analysis of top hits reveals the EZH2 interactome (right). (B) Endogenous TRIM28 in A375 cells was immunoprecipitated and immunoblotted to detect EZH2. (C) Western blot validating shRNA-mediated knockdown of TRIM28 in A375 cells (shTRIM28). (D) qPCR analysis revealing that SULF1 mRNA expression is decreased upon stable knockdown of TRIM28 . (E) Number of DEGs in the RNA-seq data (FDR p-value ≤ 0.01, |FC| ≥ 2). The number of significantly up- and down-regulated genes for shEZH2 and shTRIM28 is included in the inset table. (F) Venn diagram showing overlap of up- and down-regulated genes in shEZH2 and shTRIM28 RNA-seq datasets (FDR p-value ≤ 0.01, |FC| ≥ 2). (G) Pathway enrichment analysis of overlapping DEGs between shEZH2 and shTRIM28 populations (FDR p-value ≤ 0.01, |FC| ≥ 2). (H) Heatmap of expression z-scores for a subset of co-activated DEGs shared between shEZH2 and shTRIM28 cells relative to shCTRL controls. Red represents higher expression and blue indicates lower expression. Data are presented as mean ± SD (n = 3 independent biological replicates, n = 4 independent biological replicates for RNA-seq experiments), ***p<0.001, **p<0.01, *p<0.05 by two-sided t-test.

Article Snippet: Membranes were probed with primary antibodies against EZH2 (Cell Signaling Technology, #5246; 1:1000), EZH2 (Active Motif, #39076, 1:200), TRIM28 (Cell Signaling Technology, #85322; 1:1000), SUZ12 (Cell Signaling Technology, #3737; 1:1000), EED (Cell Signaling Technology, #85322; 1:1000), H3K27me3 (Cell Signaling Technology, #9733; 1:1000), Histone H3 (Cell Signaling Technology, #3638; 1:1000), or β-actin (Cell Signaling Technology, #3700; 1:5000) at 4°C overnight.

Techniques: Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Western Blot, shRNA, Knockdown, Expressing, RNA Sequencing

Journal: bioRxiv

Article Title: A non-canonical EZH2/TRIM28 epigenetic axis drives heparan sulfate remodeling and melanoma metastasis

doi: 10.64898/2026.01.21.700969

Figure Lengend Snippet: Profile plots showing genome-wide localization of (A) TRIM28, (B) EED, and (C) H3K27me3 relative to annotated gene bodies in A375 wild-type cells (hg38 genome, TSS: transcription start site, TES: transcription end site). Genome browser tracks showing TRIM28, EED, and H3K27me3 ChIP-seq signal at the SULF2 (D) and SULF1 (E) loci. (F) Venn diagram showing overlap between genes positively regulated by EZH2 and TRIM28 (RNA-seq) and genes bound by TRIM28 (ChIP-seq). (G) ChIP–qPCR assessing TRIM28-dependent enrichment of EZH2 at the SULF1 locus in control and TRIM28-depleted A375 melanoma cells. Data are presented as mean ± SD (n = 3 independent biological replicates), ***p<0.001, **p<0.01, *p<0.05 by two-sided t-test.

Article Snippet: Membranes were probed with primary antibodies against EZH2 (Cell Signaling Technology, #5246; 1:1000), EZH2 (Active Motif, #39076, 1:200), TRIM28 (Cell Signaling Technology, #85322; 1:1000), SUZ12 (Cell Signaling Technology, #3737; 1:1000), EED (Cell Signaling Technology, #85322; 1:1000), H3K27me3 (Cell Signaling Technology, #9733; 1:1000), Histone H3 (Cell Signaling Technology, #3638; 1:1000), or β-actin (Cell Signaling Technology, #3700; 1:5000) at 4°C overnight.

Techniques: Genome Wide, ChIP-sequencing, RNA Sequencing, ChIP-qPCR, Control

Journal: bioRxiv

Article Title: A non-canonical EZH2/TRIM28 epigenetic axis drives heparan sulfate remodeling and melanoma metastasis

doi: 10.64898/2026.01.21.700969

Figure Lengend Snippet: (A) Study design of an A375 melanoma xenograft model in NSG mice to assess primary tumor growth and metastatic dissemination of SULF1-deficient cells. (B) Total primary tumor volume measured longitudinally (mm³). Mice were euthanized upon reaching a humane endpoint defined as total tumor burden ≥ 2000 mm³, followed by excision of primary tumors and collection of organs for metastatic analysis (n = 10 mice per group; data are presented as mean ± SEM and represent total tumor volume per mouse, p = 0.0022). (C) Contingency analysis showing the number of mice with detectable metastases, as assessed by ex vivo bioluminescence imaging, in any of the indicated organs (lungs, liver, brain, bone, or lymph nodes) (n = 10 mice per group). (D) Contingency plot depicting the number of mice with observed metastases in the lung. Inset: representative image of a shCTRL-injected mouse lung showing luminescence signal from A375 cells is included as an inset. (E-F) Quantification of tumor necrosis expressed as necrotic area (red) relative to total tumor area, determined by H&E staining and QuPath and confirmed by a pathologist (n = 10 mice per group, data represent an average of total tumor necrotic area per mouse). (G) RNA-seq data from the TCGA-SKCM cohort comparing EZH2 , TRIM28 , and SULF1 expression in primary melanoma tumors (n = 103) versus metastatic melanoma tumors (n = 369). Data are presented as mean ± SEM, ***p<0.001, **p<0.01, *p<0.05, unless otherwise stated above. Statistical significance was determined using two-sided tests as indicated (tumor growth: mixed-effects model for repeated measures; metastasis incidence: chi-squared test; tumor necrosis: two-sided t-test).

Article Snippet: Membranes were probed with primary antibodies against EZH2 (Cell Signaling Technology, #5246; 1:1000), EZH2 (Active Motif, #39076, 1:200), TRIM28 (Cell Signaling Technology, #85322; 1:1000), SUZ12 (Cell Signaling Technology, #3737; 1:1000), EED (Cell Signaling Technology, #85322; 1:1000), H3K27me3 (Cell Signaling Technology, #9733; 1:1000), Histone H3 (Cell Signaling Technology, #3638; 1:1000), or β-actin (Cell Signaling Technology, #3700; 1:5000) at 4°C overnight.

Techniques: Ex Vivo, Imaging, Injection, Staining, RNA Sequencing, Expressing