Journal: Pharmaceutics

Article Title: Novel Dimer Derivatives of PF-543 as Potential Antitumor Agents for the Treatment of Non-Small Cell Lung Cancer

doi: 10.3390/pharmaceutics14102035

Figure Lengend Snippet: Effects of PF-543 and PF-543 derivatives on A549 cells biological result; ( a ) Activities of SK1 and SK2 at 10 μM concentrations of PF-543 and PF-543 derivatives; ( b ) Protein was extracted and detected SK1, SK2, and β-actin protein levels by Western blotting. Each protein level was normalized by β-actin. Western blot was performed in three independent experiments; ( c ) The mRNA levels of SK1 and SK2 were quantified by qRT-PCR by treating A549 cells with PF-543 or PF-543 derivatives at 5 and 10 μM for 24 h. SK1 and SK2 mRNA was normalized to cyclophilin; ( d ) Sphingosine, ceramide, and S1P levels were measured in A549 cells with 5 and 10 μM PF-543 and PF-543 derivatives. The data are expressed as the mean ± S.D. * p < 0.05, *** p < 0.001 compared with control group.

Article Snippet: After blocking with 5% BSA, the slides were incubated overnight at 4 °C with KI67 (Cell Signaling Technology, Danvers, MA, USA), Caspase-3 (Abcam, Cambridge, UK), and S1P antibodies (Echelon Biosciences inc, Salt Lake City, UT).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Pharmaceutics

Article Title: Novel Dimer Derivatives of PF-543 as Potential Antitumor Agents for the Treatment of Non-Small Cell Lung Cancer

doi: 10.3390/pharmaceutics14102035

Figure Lengend Snippet: Compound 4 inhibits the growth of A549 cell-induced tumor xenograft in nude Balb/c mice; ( a ) The effectiveness of inhibiting tumor growth was observed in A549 tumor-bearing nude Balb/c mice after intraperitoneal administration of vehicle or Compound 4 (5 mg/kg) 3 times a week for a period of 29 days; ( b ) Photographs were taken and weights were measured for isolated tumors from each group on day 29 after treatment; ( c – e ) Immunohistochemistry of tumor sections was performed for KI67, caspase-3 and S1P (scale bar 10 μm, magnification 200X). Data are expressed as the mean ± S.D. * p < 0.05, and ** p < 0.01, compared with control group.

Article Snippet: After blocking with 5% BSA, the slides were incubated overnight at 4 °C with KI67 (Cell Signaling Technology, Danvers, MA, USA), Caspase-3 (Abcam, Cambridge, UK), and S1P antibodies (Echelon Biosciences inc, Salt Lake City, UT).

Techniques: Isolation, Immunohistochemistry

Journal: Pharmaceutics

Article Title: Novel Dimer Derivatives of PF-543 as Potential Antitumor Agents for the Treatment of Non-Small Cell Lung Cancer

doi: 10.3390/pharmaceutics14102035

Figure Lengend Snippet: Change of blood chemistry in tumor xenograft mice for 29 days treated with vehicle and Compound 4 .

Article Snippet: After blocking with 5% BSA, the slides were incubated overnight at 4 °C with KI67 (Cell Signaling Technology, Danvers, MA, USA), Caspase-3 (Abcam, Cambridge, UK), and S1P antibodies (Echelon Biosciences inc, Salt Lake City, UT).

Techniques:

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: Expressing, Chromatin Immunoprecipitation

Journal: JCI Insight

Article Title: Linking epigenetic dysregulation, mitochondrial impairment, and metabolic dysfunction in SBMA motor neurons

doi: 10.1172/jci.insight.136539

Figure Lengend Snippet: Flow diagram depicting low acetyl-CoA levels as a central metabolic change that connects reduced expression of metabolic genes to mitochondrial ATP production impairment. From left, altered p300/CBP activity, which may be due to sequestration and depletion by the mutant AR (40), leads to reduced H3K27ac. This in turn contributes to repression of ETC and metabolic genes. Repression of these genes perturbs compensatory metabolic pathways, which results in insufficient production of acetyl-CoA. Acetyl-CoA is a common substrate for both the ETC function and p300/CBP HAT activity. Low acetyl-CoA (substrate) contributes to low ATP production and mitochondrial dysfunction. In parallel, when the energy demands of the cell increase, mitochondria respond by consuming more acetyl-CoA (substrate). The reduction of acetyl-CoA contributes to repression of ETC and metabolic genes through impaired p300/CBP acetylation.

Article Snippet: Membranes were probed with 1:1000 rabbit anti-AR (Santa Cruz Biotechnology Inc., sc-13062), 1:5000 mouse anti–α-tubulin (MilliporeSigma, T6199), 1:1000 mouse anti-histone H1 (Thermo Fisher Scientific, PA5-30055), 1:1000 rabbit anti–ac-P300/CBP (Cell Signaling Technology, 4771), 1:1000 rabbit anti-P300 (Cell Signaling Technology, 86377), 1:1000 mouse anti-HDAC1 (Cell Signaling Technology, 5356), 1;1000 rabbit anti-H3 (MilliporeSigma, 05-928), 1:1000 rabbit anti–cleaved caspase-3 (Cell Signaling Technology, 9661), 1:1000 rabbit anti–caspase-3 (Cell Signaling Technology, 9662), 1:5000 mouse anti-actin (MilliporeSigma, A2228), and 1:1000 rabbit anti-vinculin (Cell Signaling Technology, 13901).

Techniques: Expressing, Activity Assay, Mutagenesis