116 research outputs found

    Representative pattern of AcH4 expression in spermatids treated with 50 µM Curcumin for 48

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    <p> <b>h.</b> (A). Immunostaining of AcH4 in spermatids. Step: Developmental steps of spermiogensis. Red: Signals of AcH4. Green: Acrosomes highlighted with lectin PNA. Blue: Nuclei counterstained by Hoechst 33342. Bars = 5 µm. The quantitative analysis of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048673#pone-0048673-g006" target="_blank">Figure 6</a> was listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048673#pone.0048673.s005" target="_blank">Table S3</a>. (B). Immunoblot of AcH4 in spermatids.</p

    Histone Acetylase Inhibitor Curcumin Impairs Mouse Spermiogenesis–An <em>In Vitro</em> Study

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    <div><p>In the previous study, we unraveled the unique “erasure strategy” during the mouse spermiogenesis. Chromatin associated proteins sequentially disassociated from the spermatid chromosome, which led to the termination of transcription in elongating spermatids. By this process, a relatively naïve paternal chromatin was generated, which might be essential for the zygotic development. We supposed the regulation of histone acetylation played an important role throughout this “erasure” process. In order to verify this hypothesis, we treated mouse spermatids <em>in vitro</em> by histone acetylase (HAT) inhibitor Curcumin. Our results showed an inhibiting effect of Curcumin on the growth of germ cell line in a dose-dependent manner. Accordingly, the apoptosis of primary haploid spermtids was increased by Curcumin treatment. As expected, the acetylated histone level was downregulated. Furthermore, we found the transcription in spermatids ceased in advance, the dynamics of chromatin associated factors was disturbed by Curcumin treatment. The regulation of histone acetylation should be one of the core reprogramming mechanisms during the spermiogenesis. The reproductive toxicity of Curcumin needs to be thoroughly investigated, which is crucial for its further clinical application.</p> </div

    A working hypothesis for the molecular mechanisms of reprogramming in mouse spermiogenesis.

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    <p>(A) During normal spermiogenesis, transcription restarted in spermatids specific HATs were generated. Later, HDACs catalyzed the deacetylation of histones, leading to the CAFs dissociation and transcription cessation. In the meanwhile, the gradually accumulated specific HATs guided the degradation of HDACs, in that case they could induce the histone hyperacetylation in elongating spermatids.The AcH signal then recruited remodeling factors to execute the histone substitution and nuclear condensation. (B) When spermatids were treated with Curcumin, hypoacetylation was induced, resulting in a premature erasure procedure of CAFs and transcription. The spermatid specific HATs were missing, whereas the HDACs retained. At last, the transformation of spermatids was impaired. A: acetylated histones; s-HAT: spermatid specific histone acetylase; HDAC: histone deacetylase; TAFs: transcription associated factors; TRs: transcription regulators; RFs: remodeling factors; EPs: epigenetic modifiers; TPs: transition proteins. (This figure was modified from Tsankova et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048673#pone.0048673-Tsankova1" target="_blank">[40]</a>).</p

    One representative assay of haploid spermatids aggregating by FACS.

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    <p>(A). Hoechst33342 stained primary testicular cells were analyzed in SORP FACSAria II FACScan flow cytometer. <b>P4</b> indicated the haploid cell population. (B). The gathered haploid spermatids were confirmed by DNA content analysis (lower) with the total testicular cells used as a control (upper).</p

    Transcription was affected by 50 µM Curcumin treatment.

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    <p>(A). Quantitative RT-PCR results of mRNA levels of tested genes. Spermatids were incubated with 50 µM Curcumin for 3 h before qPCR executed (Mean ± SD, n = 3). #<i>p<</i>0.05. (B). Quantitative RT-PCR results of mRNA levels of tested genes. Spermatids were incubated with 50 µM Curcumin for 48 h before qPCR executed (Mean ± SD, n = 3). #<i>p<</i>0.05. *<i>p<</i>0.01. (C). Global transcription status illustrated by <i>in vitro</i> run-on assay. Step: Developmental steps of spermiogenesis. Red: Signals of BrUTP incorporation. Green: Acrosomes highlighted with lectin PNA. Blue: Nuclei counterstained by Hoechst 33342. Bars = 5 µm.</p

    Apoptosis analysis of male germ cells after Curcumin treatment.

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    <p>(A). Apoptosis of testicular cells treated with 50 µM Curcumin for 3 h (Mean ± SD, n = 3). (B). Apoptosis of spermatids treated with 50 µM Curcumin for 3 h (Mean ± SD, n = 3). #<i>p<</i>0.05. (C). Apoptosis of testicular cells treated with 50 µM Curcumin for 48 h (Mean ± SD, n = 4). #<i>p<</i>0.05.</p

    Representative patterns of chromatin associated factors expression in spermatids treated with 50 µM Curcumin for 48

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    <p> <b>h.</b> (A). Immunostaining of TBP, TAF1, AP2α, TOPOIIβ, H3K4Me3 and H4K20Me3 in Curcumin-treated spermatids. Red: Signals of given target. Green: Acrosomes highlighted with lectin PNA. Blue: Nuclei counterstained by Hoechst 33342. Bars = 5 µm. (B). Immunostaining of Hdac1 in Curcumin-treated spermatids. Red: Signals of given target. Green: Acrosomes highlighted with lectin PNA. Blue: Nuclei counterstained by Hoechst 33342. Bars = 5 µm. The quantitative analysis of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048673#pone-0048673-g006" target="_blank">Figure 6</a> was listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048673#pone.0048673.s005" target="_blank">Table S3</a>.</p

    Table_1_Identification of immune cell function in breast cancer by integrating multiple single-cell data.xlsx

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    Breast cancer has now become the most commonly diagnosed cancer worldwide. It is a highly complex and heterogeneous disease that comprises distinct histological features and treatment response. With the development of molecular biology and immunology, immunotherapy has become a new field of breast cancer treatment. Identifying cell-type-specific genes critical to the immune microenvironment contributes to breast cancer treatment. Single-cell RNA sequencing (scRNA-seq) technology could serve as a powerful tool to analyze cellular genetic information at single-cell resolution and to uncover the gene expression status of each cell, thus allowing comprehensive assessment of intercellular heterogeneity. Because of the influence of sample size and sequencing depth, the specificity of genes in different cell types for breast cancer cannot be fully revealed. Therefore, the present study integrated two public breast cancer scRNA-seq datasets aiming to investigate the functions of different type of immune cells in tumor microenvironment. We identified total five significant differential expressed genes of B cells, T cells and macrophage and explored their functions and immune mechanisms in breast cancer. Finally, we performed functional annotation analyses using the top fifteen differentially expressed genes in each immune cell type to discover the immune-related pathways and gene ontology (GO) terms.</p

    Mechanical analysis for FPP under different control conditions.

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    <p>(a) Δσ=0 MPa, T<sub><b>0</b></sub>=3.6 MPa; (b) Δσ=0 MPa, T<sub><b>0</b></sub>=4.2 MPa; (c) Δσ=5 MPa, T<sub><b>0</b></sub>=3.6 MPa; (d) Δσ=5 MPa, T<sub><b>0</b></sub>=4.2 MPa; (e) Δσ=10 MPa, T<sub><b>0</b></sub>=3.6 MPa; (f) Δσ=10 MPa, T<sub><b>0</b></sub>=4.2 MPa.</p

    Variation rule of branch fracture propagation pressure.

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    <p>(a) Δσ=0 MPa, K<sub><b>IC</b></sub>=0.8 MPa m<sup>0.5</sup>; (b) Δσ=0 MPa, K<sub><b>IC</b></sub>=1.6 MPa m<sup>0.5</sup>; (c) Δσ=5 MPa, K<sub><b>IC</b></sub>=0.8 MPa m<sup>0.5</sup>; (d) Δσ=5 MPa, K<sub><b>IC</b></sub>=1.6 MPa m<sup>0.5</sup>; (e) Δσ=10 MPa, K<sub><b>IC</b></sub>=0.8 MPa m<sup>0.5</sup>; (f) Δσ=10 MPa, K<sub><b>IC</b></sub>=1.6 MPa m<sup>0.5</sup>.</p
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