Journal: Biomedicines

Article Title: R -α-Lipoic Acid and 4-Phenylbutyric Acid Have Distinct Hypolipidemic Mechanisms in Hepatic Cells

doi: 10.3390/biomedicines8080289

Figure Lengend Snippet: PBA enhances the acetylation of CBP/p300 and histone H3 at the CPT1A and INSIG2 promoters in Huh7-shTSC2 cells. PBA (8 mM, 6 h) induced the gene expression of CPT1A and INSIG2 ( A ), and the acetylation status of histone H3 and histone H4 ( B ). Chromatin immunoprecipitation (ChIP) assay revealed PBA-mediated enrichment of acetyl CBP/p300 (Ac-p300) and acetyl histone H3 (Ac-H3) at the CPT1A and INSIG2 promoters ( C ). Statistical significance was determined using Student’s t -test, * p < 0.05, n = 4.

Article Snippet: After pre-clearing the sheared chromatin, samples were subjected to immunoprecipitation using the following the antibodies: anti-acetyl-CBP [Lys1535]/p300 [Lys1499] (Cell Signaling Technology #4771S, Danvers, MA, USA), anti-acetyl-histone H3 (Millipore #06–599, Burlington, MA, USA), anti-histone H3 (Millipore #07–690).

Techniques: Gene Expression, Chromatin Immunoprecipitation

Journal: Nature Communications

Article Title: Assay interference and off-target liabilities of reported histone acetyltransferase inhibitors

doi: 10.1038/s41467-017-01657-3

Figure Lengend Snippet: Reported HAT inhibitors cause nonspecific cell assay readouts. a Double p300 and CBP knockdowns do not affect cell viability after 2 d. Data are expressed as mean ± SEM of three technical replicates (ns, not significant; two-tailed Student’s t -test). b Double p300 and CBP knockdowns efficiently reduce H3K27ac levels in HEK293T and MCF7 cells. Molecular weight of probed protein is indicated in kilodaltons (kDa) as verified by molecular weight markers. Data are representative results from one of two independent experiments. c Many reported HAT inhibitors exert anti-proliferative effects at low micromolar compound concentrations in (top) MCF7 and (bottom) HEK293T cells as monitored by cell confluence 24 h after addition of compound. Note the nonspecific interference compounds NSC-663284 (N), rottlerin (R), and 24 – 27 (chemical structures shown as inset) demonstrate similar anti-proliferative effects. C, DMSO control. Red error bars, statistically significant difference from respective DMSO controls ( p < 0.05; two-tailed Student’s t -test and Holm–Sidak method). Data are expressed as mean ± SD of three technical replicates and are representative results from one of two independent experiments. d Many reported HAT inhibitors also cause nonspecific changes in H3K27ac and p300 levels in cells at low micromolar compound concentrations. Molecular weights of protein analytes are indicated in kDa as verified by molecular weight markers (Supplementary Fig. ). Note the same known interference compounds from panel c can also decrease H3K27ac levels. Data are representative results from one of two independent experiments

Article Snippet: Blots were blocked for 1 h in blocking buffer (5% milk in 0.1% Tween 20/PBS) and incubated with respective primary antibodies: H3K27ac (1:1000, CST #8173), H3 (1:1000, Abcam #10799), p300 (1:2000, Bethyl #A300-358A), CBP (1:1000, Bethyl #A300-362A), actin (1:3000, Abcam #3280), α-tubulin (1:1000, 12G10 DHSB), and α-tubulin-K40ac (1:2000, Abcam #179484) in blocking buffer overnight at 4 °C.

Techniques: Two Tailed Test, Molecular Weight

Journal: Medicine

Article Title: Pathological discrimination between luteinized thecoma associated with sclerosing peritonitis and thecoma

doi: 10.1097/MD.0000000000033911

Figure Lengend Snippet: Comparison of molecular pathological markers in luteinized thecoma associated with sclerosing peritonitis and thecoma.

Article Snippet: The tissue sections were stained for MGAT5B (1:200; catalog number 16993; Proteintech, Wuhan, China), NCOA3 (1:50, catalog number 20032, Proteintech), Vimentin (1:200, catalog number 10366, Proteintech), Catenin beta-1 (β-Catenin; 1:200, catalog number 17565, Proteintech), receptor tyrosine-protein kinase erbB-2 (HER2; 1:600, catalog number 18299, Proteintech), proliferation marker protein Ki-67 (MKI67; 1:5000, catalog number 27309, Proteintech), estrogen receptor (ESR; 1:200, catalog number 21244, Proteintech), progesterone receptor (PGR; 1:200, catalog number 25871, Proteintech), CD99 antigen (CD99; 1:200; catalog number abs136282; Absin, Shanghai, China), Wilms tumor protein (WT1; 1:200, catalog number abs135851, Absin), and hematoxylin-eosin as follows.

Techniques: Comparison

Journal: Medicine

Article Title: Pathological discrimination between luteinized thecoma associated with sclerosing peritonitis and thecoma

doi: 10.1097/MD.0000000000033911

Figure Lengend Snippet: Molecular pathological markers show significant ascendant expression in LTSP. (A–F) Immunohistochemical staining of alpha-1,6-mannosylglycoprotein 6-beta- n -acetylglucosaminyltransferase B (MGAT5B), nuclear receptor coactivator 3 (NCOA3) and proliferation marker protein Ki-67 (MKI67) are respectively (A, C, E) diffusely positive in the luteinized cells but (B, D, F) weak to moderate in the thecoma cells. (G) Statistical analysis of the 3 markers. Data are expressed as mean ± SD. n = 9 to 11 sections per marker (G, LTSP); n = 81 to 85 sections per marker (G, Thecoma). *** P < .001 ( t test). Scale bar, 500 μm. AOD = average optical density, LTSP = luteinized thecoma associated with sclerosing peritonitis.

Article Snippet: The tissue sections were stained for MGAT5B (1:200; catalog number 16993; Proteintech, Wuhan, China), NCOA3 (1:50, catalog number 20032, Proteintech), Vimentin (1:200, catalog number 10366, Proteintech), Catenin beta-1 (β-Catenin; 1:200, catalog number 17565, Proteintech), receptor tyrosine-protein kinase erbB-2 (HER2; 1:600, catalog number 18299, Proteintech), proliferation marker protein Ki-67 (MKI67; 1:5000, catalog number 27309, Proteintech), estrogen receptor (ESR; 1:200, catalog number 21244, Proteintech), progesterone receptor (PGR; 1:200, catalog number 25871, Proteintech), CD99 antigen (CD99; 1:200; catalog number abs136282; Absin, Shanghai, China), Wilms tumor protein (WT1; 1:200, catalog number abs135851, Absin), and hematoxylin-eosin as follows.

Techniques: Expressing, Immunohistochemical staining, Staining, Marker

Journal: Medicine

Article Title: Pathological discrimination between luteinized thecoma associated with sclerosing peritonitis and thecoma

doi: 10.1097/MD.0000000000033911

Figure Lengend Snippet: MGAT5B-NCOA3 fusion gene in LTSP of the ovary. Schematic representation of the MGAT5B-NCOA3 fusion transcript including the exons and bases sequence involved. The breakpoint of the 5′ and 3′ partner genes are represented as red dotted lines. LTSP = luteinized thecoma associated with sclerosing peritonitis, MGAT5B = alpha-1,6-mannosylglycoprotein 6-beta- n -acetylglucosaminyltransferase B, NCOA3 = nuclear receptor coactivator 3.

Article Snippet: The tissue sections were stained for MGAT5B (1:200; catalog number 16993; Proteintech, Wuhan, China), NCOA3 (1:50, catalog number 20032, Proteintech), Vimentin (1:200, catalog number 10366, Proteintech), Catenin beta-1 (β-Catenin; 1:200, catalog number 17565, Proteintech), receptor tyrosine-protein kinase erbB-2 (HER2; 1:600, catalog number 18299, Proteintech), proliferation marker protein Ki-67 (MKI67; 1:5000, catalog number 27309, Proteintech), estrogen receptor (ESR; 1:200, catalog number 21244, Proteintech), progesterone receptor (PGR; 1:200, catalog number 25871, Proteintech), CD99 antigen (CD99; 1:200; catalog number abs136282; Absin, Shanghai, China), Wilms tumor protein (WT1; 1:200, catalog number abs135851, Absin), and hematoxylin-eosin as follows.

Techniques: Sequencing

Journal: Medicine

Article Title: Pathological discrimination between luteinized thecoma associated with sclerosing peritonitis and thecoma

doi: 10.1097/MD.0000000000033911

Figure Lengend Snippet: MGAT5B-NCOA3 fusion gene is most common in LTSP. (A–C) MGAT5B-NCOA3 fusion gene is most common in (A) luteinized cells, but (B) rare in thecoma cells and (C) normal fimbriae tubae cells as assessed by FISH. FISH showing fusion signals for MGAT5B-NCOA3 with the overlapping yellow fluorescent signal (white arrows: orange, NCOA3; green, MGAT5B; yellow, fusion area). (D) Statistical analysis of positive incidence. Data are expressed as mean ± SD percentage. n = 7 cases (D, LTSP); n = 5 cases (D, Thecoma); n = 2 cases (D, Control). *** P < .001 (one-way analysis of variance and post hoc test). Scale bar, 250 μm. FISH = fluorescence in situ hybridization, LTSP = luteinized thecoma associated with sclerosing peritonitis, MGAT5B = alpha-1,6-mannosylglycoprotein 6-beta- n -acetylglucosaminyltransferase B, NCOA3 = nuclear receptor coactivator 3, ns = no significance.

Article Snippet: The tissue sections were stained for MGAT5B (1:200; catalog number 16993; Proteintech, Wuhan, China), NCOA3 (1:50, catalog number 20032, Proteintech), Vimentin (1:200, catalog number 10366, Proteintech), Catenin beta-1 (β-Catenin; 1:200, catalog number 17565, Proteintech), receptor tyrosine-protein kinase erbB-2 (HER2; 1:600, catalog number 18299, Proteintech), proliferation marker protein Ki-67 (MKI67; 1:5000, catalog number 27309, Proteintech), estrogen receptor (ESR; 1:200, catalog number 21244, Proteintech), progesterone receptor (PGR; 1:200, catalog number 25871, Proteintech), CD99 antigen (CD99; 1:200; catalog number abs136282; Absin, Shanghai, China), Wilms tumor protein (WT1; 1:200, catalog number abs135851, Absin), and hematoxylin-eosin as follows.

Techniques: Control, Fluorescence, In Situ Hybridization

Journal: Frontiers in Neurology

Article Title: Identification of m1A/m6A/m5C/m7G-related genes and clusters associated with neuropathic pain

doi: 10.3389/fneur.2026.1592545

Figure Lengend Snippet: Validation of expression of the six m1A/m6A/m5C/m7G-related DEGs ( FTO , METTL3 , NSUN2 , YTHDF3 , WDR4 , and EIF4E ) in rats after 3, 7, and 14 days of SNL surgery. n = 3 per group. SNL, spinal nerve ligation. (A) qPCR results. (B) Western blot analysis results. * p < 0.05.

Article Snippet: After blocking, the membranes were incubated with the following primary antibodies: Nsun2 (1:1000, PH6626, ab-mart, Shanghai, China), Mettl3 (1:1000, 15,073-1-AP, Proteintech, Rosemont, IL, United States), Ythdf3 (1:500, 25,537-1-AP, Proteintech), FTO (1:1000, PA2776, ab-mart); Wdr4 (1:1000, PS17092, ab-mart), Eif4e (1:1000, 11,149-1-AP, Proteintech), and GAPDH (1:20000, 10,494-1-AP, Proteintech) overnight at 4 °C.

Techniques: Biomarker Discovery, Expressing, Ligation, Western Blot