Journal: Molecular cell

Article Title: KAP1 is a Chromatin Reader that Couples Steps of RNA Polymerase II Transcription to Sustain Oncogenic Programs

doi: 10.1016/j.molcel.2020.04.024

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Histone H4 N-terminal peptide, biotinylated (1–23) , EpiCypher , Cat# 12-0029.

Techniques: Virus, Mutagenesis, Recombinant, Membrane, Protease Inhibitor, SYBR Green Assay, Western Blot, Staining, Plasmid Preparation, RNA HS Assay, Reverse Transcription, Bicinchoninic Acid Protein Assay, Cloning, shRNA, Software

Journal: The Journal of Headache and Pain

Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

doi: 10.1186/s10194-026-02328-9

Figure Lengend Snippet: S100A11 knockdown alleviates microglial inflammation by inhibiting mitophagy through the TFEB/H3K27ac axis. A-B Western blot analysis of TFEB, TFEC, and TFE3 protein expression levels in the cytoplasm and nucleus of primary microglia, along with the corresponding detection and quantification plots ( D-F , for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). C Immunofluorescence staining of TFEB in primary microglia. G Western blot analysis of pink1, Parkin, P62, LC3II protein levels of primary microglia; H-K Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). M Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panel L . and relative fluorescence intensity was quantified in panel L ( n = 3). N Western blot analysis of H4K16ac, H4K12ac, H3K27ac, H3K18ac, H3K9ac protein levels of microglia; O-S Relative levels normalized to histone H3 were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). T Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; U-X Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Z Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green), and relative fluorescence intensity was quantified in panel Y ( n = 3). Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

Article Snippet: Membranes were blocked with 5% skim milk for 60 min, rinsed with TBST, and incubated with primary antibodies at 4 °C overnight (β-actin: Proteintech, 66009-1-Ig, 1:10000; S100A11: Abcam, ab169530, 1:1000; NLRP3: AdipoGen, AG-20B-0014, 1:1000; GSDMD-N: Abcam, ab215203, 1:1000; ASC: Wanleibio, WL02462, 1:500; Caspase1: CST, 89332, 1:1000; Bax: Proteintech, 50599-2-Ig, 1:2000; Bcl-2: abclone, A19693, 1:2000; COXIV: Proteintech, 11242-1-AP, 1:1000; LC3: Proteintech, 14600-1-AP, 1:2000; P62: Proteintech, 18420-1-AP, 1:2000; Parkin: Proteintech, 14060-1-AP, 1:1000; Pink1: Proteintech, 23274-1-AP, 1:2000; H3: Proteintech, 17168-1-AP, 1:2000; TFEB: Proteintech, 13372-1-AP, 1:1000; TFEC: Proteintech, 13547-1-AP, 1:1000; TFE3: Proteintech, 14480-1-AP, 1:1000; H3K9ac: CST, 9649, 1:1000; H3K18ac: CST, 13998, 1:1000; H3K27ac: CST, 8173, 1:1000; H4K12ac; CST, 13944, 1:1000; H4K16ac: CST, 13534, 1:1000).

Techniques: Knockdown, Western Blot, Expressing, Immunofluorescence, Staining, Labeling, Fluorescence

Journal: PLoS ONE

Article Title: Astaxanthin Inhibits JAK/STAT-3 Signaling to Abrogate Cell Proliferation, Invasion and Angiogenesis in a Hamster Model of Oral Cancer

doi: 10.1371/journal.pone.0109114

Figure Lengend Snippet: (mean ± SD; n = 3). A. Transcript expression level of JAK-2 and STAT-3 in various experimental groups as determined by kinetic PCR. Data are the mean ± SD of three independent experiments. ♣ p<0.05 versus control. *p<0.05 versus DMBA. B. Representative immunoblot analysis. Protein samples (100 μg/lane) resolved on SDS-PAGE were probed with corresponding antibodies. GAPDH was used as loading control for cytosol and whole tissue homogenates. Histone H2B was used as loading control for nuclear proteins. Phosphorylated proteins are normalized by their unphosphorylated form. C. Densitometric analysis. The protein expression from control lysates for three determinations was designated as 100% in the graph. Each bar represents the protein expression of three determinations. ♣ p<0.05 versus control. *p<0.05 versus DMBA. D. Levels of pSTAT tyr705 (ELISA). E. Representative photomicrographs of immunohistochemical staining of pSTAT-3 in control and experimental animals (20X).

Article Snippet: Antibodies for IL-6, GAPDH, Cyclin D1, PCNA, p21, MMP-2, MMP-9, TIMP-2, RECK, VEGF, VEGFR2, HIF1α, were purchased from Santa Cruz Biotechnology, USA. pJAK-2 tyr1007/1008 , JAK-2, pSTAT-3 tyr705 , STAT-3 and histone (H2B) antibodies and BrdU, STAT-3 tyr705 , total cyclin D1 and pVEGFR2 tyr1175 ELISA kits were from Cell Signaling Technology, USA.

Techniques: Expressing, Control, Western Blot, SDS Page, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Astaxanthin Inhibits JAK/STAT-3 Signaling to Abrogate Cell Proliferation, Invasion and Angiogenesis in a Hamster Model of Oral Cancer

doi: 10.1371/journal.pone.0109114

Figure Lengend Snippet: (mean ± SD; n = 3). A. Transcript expression level of p21, cyclin D1 and PCNA in various experimental groups as determined by kinetic PCR. Data are the mean ± SD of three independent experiments. ♣ p<0.05 versus control. *p<0.05 versus DMBA. B. Representative immunoblot analysis. Protein samples (100 μg/lane) resolved on SDS-PAGE were probed with corresponding antibodies. GAPDH was used as loading control for cytosol and whole tissue homogenates. Histone H2B was used as loading control for nuclear proteins. C. Densitometric analysis. The protein expression from control lysates for three determinations was designated as 100% in the graph. Each bar represents the protein expression of three determinations. ♣ p<0.05 versus control. p<0.05 versus DMBA. D. Levels of total cyclin D1 (ELISA). E. Representative photomicrographs of immunohistochemical staining of PCNA in control and experimental animals (20X).

Article Snippet: Antibodies for IL-6, GAPDH, Cyclin D1, PCNA, p21, MMP-2, MMP-9, TIMP-2, RECK, VEGF, VEGFR2, HIF1α, were purchased from Santa Cruz Biotechnology, USA. pJAK-2 tyr1007/1008 , JAK-2, pSTAT-3 tyr705 , STAT-3 and histone (H2B) antibodies and BrdU, STAT-3 tyr705 , total cyclin D1 and pVEGFR2 tyr1175 ELISA kits were from Cell Signaling Technology, USA.

Techniques: Expressing, Control, Western Blot, SDS Page, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Astaxanthin Inhibits JAK/STAT-3 Signaling to Abrogate Cell Proliferation, Invasion and Angiogenesis in a Hamster Model of Oral Cancer

doi: 10.1371/journal.pone.0109114

Figure Lengend Snippet: (mean ± SD; n = 3). A. Transcript expression level of HIF-1α, VEGF and VEGFR2 in various experimental groups as determined by kinetic PCR. Data are the mean ± SD of three independent experiments. ♣ p<0.05 versus control. *p<0.05 versus DMBA. B. Representative immunoblot analysis. Protein samples (100 μg/lane) resolved on SDS-PAGE were probed with corresponding antibodies. GAPDH was used as loading control for cytosol and whole tissue homogenates. Histone H2B was used as loading control for nuclear proteins. C. Densitometric analysis. The protein expression from control lysates for three determinations was designated as 100% in the graph. Each bar represents the protein expression of three determinations. ♣ p<0.05 versus control. *p<0.05 versus DMBA. D. Levels of pVEGFR2 tyr1175 (ELISA). E. Representative photomicrographs of immunohistochemical staining of VEGF in control and experimental animals (20X).

Article Snippet: Antibodies for IL-6, GAPDH, Cyclin D1, PCNA, p21, MMP-2, MMP-9, TIMP-2, RECK, VEGF, VEGFR2, HIF1α, were purchased from Santa Cruz Biotechnology, USA. pJAK-2 tyr1007/1008 , JAK-2, pSTAT-3 tyr705 , STAT-3 and histone (H2B) antibodies and BrdU, STAT-3 tyr705 , total cyclin D1 and pVEGFR2 tyr1175 ELISA kits were from Cell Signaling Technology, USA.

Techniques: Expressing, Control, Western Blot, SDS Page, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Journal of Biological Chemistry

Article Title: The U3 Region of Moloney Murine Leukemia Virus Contains Position-independent Cis-acting Sequences Involved in the Nuclear Export of Full-length Viral Transcripts

doi: 10.1074/jbc.m113.545855

Figure Lengend Snippet: FIGURE 6. RNA analysis of full-length and internal transcripts in transfected cells. A, Western blot analysis of different fractions of cell lysates. Cell lysates of transfected cells were normalized for total protein content and subjected to SDS-PAGE. Antibodies against histone H4 and -tubulin were used. B, representative histograms show the levels of full-length and internal pLgfpSNsin RNA transcripts in the nucleus relative to those of pLgfpSNwt. **, p 0.01, ***, p 0.001. C, representative histograms show the levels of full-length and internal pLgfpSNsin RNA transcripts in the cytoplasm of transfected cells relative to those of pLgfpSNwt. **, p 0.01, ***, p 0.001. D, representative histograms show the levels of pLgfpMNsin full-length viral transcripts in the nucleus and cytoplasm of transfected cells compared with those of the parental vector (pLgfpSNwt). ***, p 0.001. RNA was extracted from the nuclear and cytoplasmic fractions of GPenvAM12 packaging cells transfected with pLgfpSNwt, pLgfpSNsin, or pLgfpMNsin. Pre-mRNA levels of the endogenous mouse GAPDH gene were determined to evaluate contamination of the cytoplasmic fractions with nuclear RNAs. Normalization of all samples was carried out by real-time RT-PCR using primers specific for 18 S rRNA. Data are expressed as the mean of at least four independent experiments S.D.

Article Snippet: Membranes were stained with antibodies against histone H4 and -tubulin (sc-25260 and sc-5286, respectively; Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Transfection, Western Blot, SDS Page, Plasmid Preparation, Quantitative RT-PCR