Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: High p53 delays melanoma formation but primarily suppresses the progression of primary to metastatic melanoma. (A) Scatter plot of age of onset of 1st pigmented lesion (PL) (n = 25 for TP-ras0/+ and n = 11 for TP-ras0/+: Mdm4+/− mice), melanomas (n = 14 for TP-ras0/+ and n = 11 for TP-ras0/+: Mdm4+/− mice), and the age of onset of all individual pigmented lesions (n = 113 for TP-ras0/+ and n = 47 for TP-ras0/+: Mdm4+/− mice). The mean is represented by the bar in the center of each plotted data series. ns, not significant (P > 0.05); 1 star, significant (P = 0.01–0.05); 2 stars, very significant (P = 0.001–0.01). (B) Histogram presenting the percentage of mice with pigmented lesions (PL) classified by tumor size and progression. The labels for each of the 5, 10, and 20 mm2 groupings refer to tumors that are ≥ the indicated size.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques:

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: High p53 prohibited tumor growth. Growth (mm2) of individual tumors of TP-ras0/+ mice (A) and TP-ras0/+: Mdm4+/− mice (B) followed every 10 days from emergence. (C) Box and whisker plot of growth rate (mm2/day) of these tumors based on histological presentation (nevus or melanoma) and size (<5, 5–10 and >10 mm2). The whiskers represent the minimum and maximum growth rates. (D) Kaplan–Meier presentation of the survival of TP-ras0/+ (n = 28) and TP-ras0/+: Mdm4+/− mice (n = 17), Log-rank test P = 0.0441.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Whisker Assay

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Histological characterization of pigmented lesions. (A) Scatter plot of p53 immunopositive cells in 25 TP-ras0/+ nevi, 12 TP-ras0/+ melanomas, 13 TP-ras0/+: Mdm4+/− nevi, and 4 TP-ras0/+: Mdm4+/− melanomas (>5 mm2) analyzed per field (0.5 mm2 area using 20× magnification). (B) Plot of tumor progression (based on size) in relation to degree of pigmentation of each pigmented lesion using the reference scale from 1 (highest, 100%) to 5 (lowest, 0–10%). A representative picture of hematoxylin and eosin (HE) sections of pigmented lesion scored by the degree of pigmentation (at the bottom).

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques:

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Effect of Nutlin-3 on clonogenecity of melanoma cell lines. (A) A representative picture (4× magnification) of colony-forming assay of four cell lines (BL: mutant p53, A04 and MM329: wild-type p53). Cell lines treated (NT) or treated with (10, 30 μM) Nutlin-3 were plated on Matrigel matrix and allowed to grow for 72 h before colony counting. All colonies ≥50 cells in the chamber were counted. (B) A graphic representation of the effect of Nutlin-3 treatment on colony counts of tested cell lines.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Mutagenesis

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Analysis of p53 target genes in melanocytes treated with Nutlin-3 for 18 h. (A) Representation of p53 target genes based on microarray results. In red, up-regulated genes; in white, genes with no differential expression; in blue, down-regulated genes. (B) Representation of up-regulated (red) and down-regulated (blue) genes in the CDKN1A network generated by Ingenuity pathway analysis tools. Arrows represent the regulatory relationships between genes. (C) Schematic representation of tumor-suppressive activity of p53 in Ras-dependent DMBA-induced progression model from a melanocyte to nevus to metastatic melanoma. GA, growth arrest.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Microarray, Expressing, Generated, Activity Assay

Journal: Scientific Reports

Article Title: Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3

doi: 10.1038/s41598-023-49694-x

Figure Lengend Snippet: Target prediction of miR-205-5p identifies HIPK3 as one of the potential targets of miR-205-5p. ( a ) The miRDB database predicted 736 potential target genes. For the TargetScan and miRWalk databases, the corresponding numbers were 590 and 4636, respectively. In total, 140 potential target genes were identified in common by all three databases. ( b ) Analysis using existing data from the GEPIA database reveals that the relative expression level of HIPK3 in CCA tissues (n = 36) is higher than in corresponding non-tumorous tissues (n = 9). ( c ) Kaplan–Meier overall survival curves according to the levels of HIPK3 expression. ( d ) Expression levels of five predicted target genes (CDK14, CREB1, HIPK3, KMT2A, and PLCB1) in the miR-205-5p inhibitor-transfected KKU-213B cell line. ( e ) Relative expression levels of HIPK3 in three different batches of KKU-213B cells transfected with miR-205-5p inhibitor. Experiments were performed in triplicate on each batch and expressed as the mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The sections were then incubated with rabbit anti-HIPK3 polyclonal antibody (1:400 dilution; cat. No. 25107-1-AP, Proteintech, USA) diluted in 5% BSA solution at 4 °C overnight.

Techniques: Expressing, Transfection

Journal: Scientific Reports

Article Title: Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3

doi: 10.1038/s41598-023-49694-x

Figure Lengend Snippet: HIPK3 might be direct target of miR-205-5p. ( a ) Relative basal expression of HIPK3 in CCA cells (KKU-055, KKU-100, and KKU-213b) and immortalized cholangiocyte cell line (MMNK1) was measured by RT-qPCR. ( b ) Relative expression of HIPK3 in the KKU-100 CCA cell line was measured at 48 h after transfection with a HIPK3 pcDNA. ( c ) Relative proliferation rates of KKU-100 after transfected with HIPK3 pcDNA were measured using MTT assay at 24, 48 and 72 h after transfection. ( d, e ) The relative cell migration rates of KKU-100 cells transfected with HIPK3 pcDNA compared with mock-transfected controls at 12, 24, 48, and 72 h were measured using wound-healing assay. ( f, g ) KKU-213B cell line was transfected with miR-205-5p inhibitor and ( h , i ) KKU-100 cell line was transfected with miR-205-5p mimic and the expression of HIPK3 was measured by Western blot analysis. Experiments were performed in triplicate for each batch and expressed as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The sections were then incubated with rabbit anti-HIPK3 polyclonal antibody (1:400 dilution; cat. No. 25107-1-AP, Proteintech, USA) diluted in 5% BSA solution at 4 °C overnight.

Techniques: Expressing, Quantitative RT-PCR, Transfection, MTT Assay, Migration, Wound Healing Assay, Western Blot

Journal: Scientific Reports

Article Title: Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3

doi: 10.1038/s41598-023-49694-x

Figure Lengend Snippet: HIPK3 immunohistochemical staining in CCA tissues (magnification, × 40). Immunohistochemistry was used to compare the H-scores of HIPK3 in normal and CCA tissues. ( a ) One entire tissue microarray slide. ( b ) The normal bile duct (NBD) expressed HIPK3 significantly more than the CCA cells. The CCA cells stained weakly positive ( c ) or strongly positive ( d ) for HIPK3 according to H-scores. ( e ) HIPK3 H-scores were compared between healthy and malignant tissues using the Mann-Whitney U test. The median H-score cutoff was 137. High expression was higher than the median (n = 44), and low expression was lower than the median (n = 44). ( f ) The overall survival time of patients with different HIPK3 expression levels (H-score) is shown by Kaplan–Meier curve with number of risk. The experiments were expressed as mean ± SD (**** p < 0.0001).

Article Snippet: The sections were then incubated with rabbit anti-HIPK3 polyclonal antibody (1:400 dilution; cat. No. 25107-1-AP, Proteintech, USA) diluted in 5% BSA solution at 4 °C overnight.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Microarray, MANN-WHITNEY, Expressing

Journal: Scientific Reports

Article Title: Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3

doi: 10.1038/s41598-023-49694-x

Figure Lengend Snippet: Association between the clinicopathological data of CCA patients and HIPK3 expression levels.

Article Snippet: The sections were then incubated with rabbit anti-HIPK3 polyclonal antibody (1:400 dilution; cat. No. 25107-1-AP, Proteintech, USA) diluted in 5% BSA solution at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3

doi: 10.1038/s41598-023-49694-x

Figure Lengend Snippet: Univariate and multivariate regression analysis of the associations between survival time and clinicopathological features of CCA patients.

Article Snippet: The sections were then incubated with rabbit anti-HIPK3 polyclonal antibody (1:400 dilution; cat. No. 25107-1-AP, Proteintech, USA) diluted in 5% BSA solution at 4 °C overnight.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: ARID1A Downregulation Predicts High PD-L1 Expression and Worse Clinical Outcome in Patients With Gallbladder Cancer

doi: 10.3389/fonc.2022.787897

Figure Lengend Snippet: Bioinformatics analysis revealed the regulation of immune-related genes and signaling pathways in AT-rich interactive domain 1A (ARID1A)-inactivated tumors. (A) Heatmap of cytotoxic T lymphocyte (CTL) signatures in tumors with high or low ARID1A expression. (B) Volcano plot presenting the downregulation of genes involved in CD8+ T-cell activation in ARID1A-inactivated tumors. (C) Gene set enrichment analysis revealed the downregulation of the immune-related pathway in ARID1A-inactivated tumors.

Article Snippet: Tissue microarray (TMA) was established in this study using formalin-fixed, paraffin-embedded surgical specimens, and the specimens were stained immunohistochemically using appropriate antibodies (anti-ARID1A [1:600, ab182560, Abcam, USA]; anti-PD-L1 [1:200, SP142, Roche, Switzerland]; anti-PD1 [1:100, ab52587, Abcam]; anti-CD8 [1:400, ab4055, Abcam]).

Techniques: Expressing, Activation Assay

Journal: Theranostics

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling

doi: 10.7150/thno.75936

Figure Lengend Snippet: HPCAL1 directly binds to RUVBL1. (A) HEK293T transfected with FLAG-HPCAL1 or control plasmid were subjected to Co-IP/MS. Co-IP, co-immunoprecipitation. MS, mass spectrometry. (B) Protein information and representative mass spectrum peak of RUVBL1. (C) HEK293T cells were co-transfected with EGFP-HPCAL1 and FLAG-RUVBL1 or RUVBL2 plasmids. Immunoblot analysis of the cell lysates and immunoprecipitates with the indicated antibodies. (D) Endogenous HPCAL1 derived from MYC-induced liver tumors was immunoprecipitated using anti-HPCAL1 antibody or isotype IgG control and subjected to immunoblot analysis using indicated antibody. (E) Scheme of HPCAL1 domain organization. EF, EF-hand. (F) HEK293T cells were co-transfected with EGFP-HPCAL1 fragment and FLAG-RUVBL1 plasmids. Immunoblot analysis of the cell lysates and immunoprecipitates with the indicated antibodies. (G) HEK293T cells were co-transfected with various EGFP-HPCAL1 mutants and FLAG-RUVBL1 plasmids. Immunoblot analysis of the cell lysates and immunoprecipitates with the indicated antibodies. (H) Scheme of RUVBL1 domain structures and Co-IP assay. HEK293T cells were co-transfected with EGFP-HPCAL1 and various FLAG-RUVBL1 fragment plasmids. Immunoblot analysis of the cell lysates and immunoprecipitates with the indicated antibodies. DI, domain I; DII, domain II; DIII, domain III. (I) Purified recombinant GST-HPCAL1 was incubated with purified his-RUVBL1. Coomassie blue staining and immunoblot analysis of the cell lysates or immunoprecipitates with indicated antibody. (J) Co-IP and immunoblot analysis of RUVBL1/HPCAL1 interaction in the presence of NaCl (10 mM), CaCl2 (10 mM), and ETGA (10 mM).

Article Snippet: For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature.

Techniques: Transfection, Control, Plasmid Preparation, Co-Immunoprecipitation Assay, Immunoprecipitation, Mass Spectrometry, Western Blot, Derivative Assay, Purification, Recombinant, Incubation, Staining

Journal: Theranostics

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling

doi: 10.7150/thno.75936

Figure Lengend Snippet: HPCAL1 was inversely associated with mTOR signaling in mice and human. (A) GSEA showed enrichment of mTOR signaling from the RNA-sequencing datasets of liver tumors of WT (n=3) and Hpcal1-/- (n=3) mice. (B) Western blotting analysis of the cell lysates from liver tumors of WT and Hpcal1-/- mice with indicated antibodies. (C) Immunoblot analysis of the cell lysates from Huh7 cells with HPCAL1 overexpression (left) or HPCAL1 depletion (right) using indicated antibodies. (D) Representative IHC images of HPCAL1 and p-mTOR of liver sections from WT and Hpcal1-deficient mice. (E) Heatmap demonstration of IHC score and correlation between PROX1, p-mTOR, p-4EBP1, SCD1, ACSS2 and RUVBL1 in HCC tissues (n=89). The criteria of IHC scoring were described in Materials and Methods. Pearson's correlation analysis was used to measure the relationship between indicated proteins in human HCC tissues. (F) Representative IHC images of indicated proteins derived from human HCC tissue microarray (TMA). (G) Kaplan-Meier curves analyses of patients with low versus high expression of indicated proteins HCC TMA. Scale bar, 50 µm.

Article Snippet: For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature.

Techniques: RNA Sequencing, Western Blot, Over Expression, Derivative Assay, Microarray, Expressing

Journal: Theranostics

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling

doi: 10.7150/thno.75936

Figure Lengend Snippet: HPCAL1 suppresses liver tumorigenesis via RUVBL1-dependent mTOR activation. (A) Immunoblot analysis of the HPCAL1-depleted Huh7 cell lysates with or without RUVBL1 knockdown. HP, HPCAL1. (B) HEK293T cells co-transfected with FLAG-RUVBL1 and HA-TEL2 or his-TTI1 in the presence or absence of HPCAL1 were subjected to Co-IP and immunoblot analysis with indicated antibodies. (C) Co-IP and immunoblot analysis of the lysates of liver tumors from WT and Hpcal1-/-. (D) Statistical analyses of the ratio of liver to body weight (upper), liver weight (middle) and the tumor number (lower) from livers of indicated mice (n-=8). (E) Representative images of gross morphology, HE and IHC staining from liver section of indicated mice. (F) Statistical analysis of the percentage of Ki67+ cells indicated mice. (G) Immunoblot analysis of the lysates of liver tumors from indicated mice using indicated antibodies. Scale bar, 50 µm.

Article Snippet: For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature.

Techniques: Activation Assay, Western Blot, Knockdown, Transfection, Co-Immunoprecipitation Assay, Immunohistochemistry

Journal: Theranostics

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling

doi: 10.7150/thno.75936

Figure Lengend Snippet: HPCAL1 suppresses liver tumorigenesis via RUVBL1-dependent mTOR activation. Short-term growth inhibition assay of HPCAL1-overexpressed (A) or HPCAL1-depleted (B) Huh7 cells treated with indicated concentration of AZD-8055. OE, overexpression. HP, HPCAL1. (C) Short-term growth inhibition assay of SCR or HPCAL1-depleted Huh7 cells transfected with SCR or siRUVBL1 and treated with indicated concentration of AZD-8055. (D) Schematic representation of MYC-Trp53 liver cancer model and mTORi treatment. This figure was drawn by using BioRender (JX24G8PWZC) (E) Statistical analyses of the ratio of liver to body weight (upper) and the number of tumors (lower) from livers of indicated mice fed with regular chow or chow with additional AZD-8055 at the dose of 67 mg/kg (n=7). (F) Representative images of gross morphology, HE and IHC staining from liver section of indicated mice. (H) Statistical analysis of the percentage of Ki67+ cells from indicated mice. (G) Kaplan-Meier curves analyses of indicated mice with or with dietary addition of AZD-8055 (n=9). Scale bar, 50 µm. *, **, *** means p < 0.05, p < 0.01, and p < 0.001.

Article Snippet: For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature.

Techniques: Activation Assay, Growth Inhibition Assay, Concentration Assay, Over Expression, Transfection, Immunohistochemistry

Journal: Cancer cell

Article Title: MUTANT EZH2 INDUCES A PRE-MALIGNANT LYMPHOMA NICHE BY REPROGRAMMING THE IMMUNE RESPONSE

doi: 10.1016/j.ccell.2020.04.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Single-cell suspensions from mouse spleens, lymph nodes and bone marrow were incubated with Fc block antibody (CD16/CD32, BD 553142) and subsequently stained using the following fluorescent-labeled anti-mouse antibodies: from eBioscience, ThermoFisher Scientific: eFluor506 anti-B220 (69–0452, dilution 1:750), APC anti-CD38 (17–0381, dilution 1:750), PE anti-CXCR4 (12–9991, dilution 1:400), APC anti-CD4 (17–0041, dilution 1:750), PerCP-Cy5.5 anti-CD45.1 (45–0453, dilution 1:750), PE-Cy7 anti-CD45.2 (25–0454, dilution 1:750), FITC anti-PD-1 (11–9985, dilution 1:200), PE-Cy7 streptavidin (25–4317, dilution 1:1000); from BD Biosciences: APC and BV786 anti-B220 (553092 and 563894, dilution 1:750), PE, PE-Cy7 and BV421 anti-FAS (554258, 557653 and 562633, dilution 1:750), FITC and BV395 anti-CD38 (558813 and 740245, dilution 1:750), PE-Cy7 and BV421 anti-CD86 (560582 and 564198, dilution 1:500), biotin anti-CXCR4 (551968, dilution 1:400), biotin anti-CXCR5 (551960, dilution 1:400), PE and BV510 anti-IgD (558597 and 563110, dilution 1:750), APC and BV421 anti-IgG1 (560089 and 562580, dilution 1:500), BUV737 anti-CD138 (564430, dilution 1:500), AF647 anti-cleaved caspase-3 (560626, dilution 1:100), BV421 anti-Ki67 (562899, dilution 1:500), BUV395 anti-BAFF-R (742871, dilution 1:400), BV650 anti-ICAM-2 (740470, dilution 1:300), BV711 anti-Ly108 (740823, dilution 1:300), AF488 anti-EZH2 (562479, dilution 1:50); from BioLegend: APC-Cy7 and PE anti-B220 (103224 and 103208, dilution 1:750), APC-Cy7 anti-CD38 (102728, dilution 1:750), PerCP-Cy5.5 anti-GL7 (144610, dilution 1:750), PerCP-Cy5.5 anti-FAS (152610, dilution 1:500), APC anti-CD86 (105012, dilution 1:500), biotin anti-CD83 (121504, dilution 1:400), PE-Cy7 and BV421 anti-CD138 (142514 and 142508, dilution 1:500), APC-Cy7 anti-CD45.2 (109824, dilution 1:750), APC streptavidin (405207, dilution 1:1000), PerCP-Cy5.5 anti-ICAM-1 (116123, dilution 1:300); from Biosearch Technologies: PE NP (N-5070–1, dilution 1:500); from Cell Signaling: AF647 anti-H3K27me3 (12158, dilution 1:400).

Techniques: Control, Blocking Assay, Recombinant, Adjuvant, Plasmid Preparation, Binding Assay, Staining, RNA Library Preparation, MicroChIP Assay, Sequencing, Microarray, Knock-In, Software, Gene Expression, Targeted Proteomics