Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Association of Filamin A expression with clinicopathological parameters

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Expressing

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing Filamin A expressivity in nonneoplastic breast tissue showing membranous and cytoplasmic positivity (×10). Right: Photomicrograph showing a membranous pattern in myoepithelial cells in fibroadenoma (×10)

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques:

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing invasive carcinoma of the breast (H and E, ×40). Right: Immunohistochemistry: Score 0 (absent) (×40) photomicrograph showing the absence of Filamin A expression in malignant cells

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Immunohistochemistry, Expressing

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing invasive carcinoma of the breast (H and E, ×40). Right: Immunohistochemistry: Score 3 (strong positivity) (×40); photomicrograph showing strong cytoplasmic positivity of Filamin A expression in malignant cells

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Immunohistochemistry, Expressing

Journal: Frontiers in Molecular Biosciences

Article Title: Long non-coding RNA lncMGC mediates the expression of TGF-β-induced genes in renal cells via nucleosome remodelers

doi: 10.3389/fmolb.2023.1204124

Figure Lengend Snippet: lncMGC-interacting proteins identified by RNA pull down followed by mass spectrometry. (A) In vitro transcribed RNAs (hlncMGC sense and antisense) were incubated with human HK2 kidney cell lysates and bound proteins isolated and analyzed by mass spectrometry (run in duplicate). 135 proteins were found to interact with sense lncMGC. (B) STRING DB ( https://string-db.org/ ) groups the proteins into RNA processing factors (such as PRPF3 and PTBP1), ribosomal proteins (such as RPL26 and RPL21), and nucleosome remodeling factors. (C) Heatmap of top 10 candidate proteins based on the number of peptides detected by MS (AS1 and AS2 refer to duplicates from antisense; Supplementary S1 and S2 refer to duplicates from sense). (D) RNA immunoprecipitation was used to validate lncMGC binding to the top 10 proteins in HK-2 cells. lncRNA NRON for IQGAP and lncTCF7 for SMARCA5 were used as positive controls ( ; ). Heatmap of the mean of three independent qPCRs is shown. Normalized affinity was calculated as the ratio of lncRNAs/positive control or as the ratio of lncRNAs/lncRNA with the highest affinity to the target protein. (E,F) The structures of SMARCA5 along with histone/DNA are shown in e (from the front) and f (from the side). Protein Data Bank, https://www.ebi.ac.uk/pdbe/entry/pdb/6ne3/analysis#assembly_1 .

Article Snippet: The cross-linked chromatin was sheared and immunoprecipitated with antibodies against H3K27Ac (Abcam, ab4729) and SMARCA5 (Boster Biol.

Techniques: Mass Spectrometry, In Vitro, Incubation, Isolation, RNA Immunoprecipitation, Binding Assay, Positive Control

Journal: Frontiers in Molecular Biosciences

Article Title: Long non-coding RNA lncMGC mediates the expression of TGF-β-induced genes in renal cells via nucleosome remodelers

doi: 10.3389/fmolb.2023.1204124

Figure Lengend Snippet: Differentially expressed candidate genes related to DKD from RNA-seq in WT and lncMGC KO5 MMC. (A) Heatmap (the mean of Log2RPKM from three independent samples) demonstrated genes associated with DKD among the DEGs. The changes in expression of Col4a3, Col4a4, Ucp2, Nox4, Nkd2 , and Pai1 were confirmed by RT-qPCR (B–G) . Effects of siSMARCA5 in WT MMC on the expressions of SMARCA5 (H) , Col4a3 (I) , Col4a4 (J) , Nkd2 (K) , Nox4 (L) , Ucp2 (M) , and Pai1 (N) . Data are shown as the mean of three independent experiments calculated from triplicate qPCRs. One-way ANOVA with post hoc Tukey’s test for multiple comparisons; ±SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.001; and **** p < 0.0001.

Article Snippet: The cross-linked chromatin was sheared and immunoprecipitated with antibodies against H3K27Ac (Abcam, ab4729) and SMARCA5 (Boster Biol.

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR

Journal: Frontiers in Molecular Biosciences

Article Title: Long non-coding RNA lncMGC mediates the expression of TGF-β-induced genes in renal cells via nucleosome remodelers

doi: 10.3389/fmolb.2023.1204124

Figure Lengend Snippet: Autoregulation of lncMGC expression. (A) H3K27ac at TSS of lncMGC shows clear increase in WT MMC treated with TGF-β but no increase in KO5 MMC under basal or TGF-β-treated conditions. ATAC-seq tracks show a decrease of ATAC-peaks at TSS of lncMGC in KO5 MMC compared to WT MMC in control and TGF-β-treated conditions. (B) Schematic of the genomic region of the promoter of mouse lncMGC/miR-379 cluster depicting Smad and CHOP binding sites and INR (TSS). Arrows indicate positions of PCR primers for Smad and CHOP sites. The arrow (INR) shows the position of INR. (C–H) H3K27ac and SMARCA5 enrichment (ChIP assays) increased by TGF-β in WT MMC at the lncMGC promoter Smad binding site and initiator (INR) site also known as the transcription start site. However, such an increase was not detected in lncMGC-KO5 MMC, suggesting lncMGC regulation of its own promoter. (I–K) Effects of siSMARCA5 on the expression of lncMGC (I) , miR-379 (J) , and H3K27ac at the Smad site of the lncMGC promoter (K) . Data are shown as the mean of three ChIP experiments calculated by triplicate qPCRs each. One-way ANOVA with post hoc Tukey’s test for multiple comparisons; ±SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.001; and **** p < 0.0001.

Article Snippet: The cross-linked chromatin was sheared and immunoprecipitated with antibodies against H3K27Ac (Abcam, ab4729) and SMARCA5 (Boster Biol.

Techniques: Expressing, Control, Binding Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Long non-coding RNA lncMGC mediates the expression of TGF-β-induced genes in renal cells via nucleosome remodelers

doi: 10.3389/fmolb.2023.1204124

Figure Lengend Snippet: Effects of lncMGC deletion and SMARCA5 siRNA (siSMARCA5) on lncMGC neighboring genes. (A) Genomic structure of the Dlk1-lncMGC region and a proposed model of regulation of neighboring genes by lncMGC RNA through SMARCA5. (B–E) Expression levels (RT-qPCR) of lncMGC neighboring genes ( Dlk1, Meg3, Rian, and Mirg ) in KO5 MMC compared to WT MMC. (F) ATAC-seq and H3K27 ChIP-seq tracks in WT and lncMGC KO5 MMC showed changes at the TSS of lncMGC in KO5 MMC compared to WT MMC in control and TGF-β-treated conditions. ATAC-peaks at the TSS of lncMGC adjacent transcripts ( Dlk1, Meg2, and Rian ) are shown in KO5 MMC compared to WT MMC in control conditions and TGF-β-treated conditions. The positions of ATAC-peaks (green) are the same as those of H3K27ac (blue), suggesting those sites are critical for the expression of these genes. (G–J) Effects of siSMARCA5 in WT MMC on Dlk1 (G) , Meg3 (H) , Rian (I) , and Mirg (J) . Data are shown as the mean of three independent experiments calculated from triplicate qPCRs. One-way ANOVA with post hoc Tukey’s test for multiple comparisons; ±SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.001; and **** p < 0.0001.

Article Snippet: The cross-linked chromatin was sheared and immunoprecipitated with antibodies against H3K27Ac (Abcam, ab4729) and SMARCA5 (Boster Biol.

Techniques: Expressing, Quantitative RT-PCR, ChIP-sequencing, Control

Journal: Frontiers in Molecular Biosciences

Article Title: Long non-coding RNA lncMGC mediates the expression of TGF-β-induced genes in renal cells via nucleosome remodelers

doi: 10.3389/fmolb.2023.1204124

Figure Lengend Snippet: Motif analyses and proposed mechanism by which lncMGC and SMARCA5 regulate DKD-related genes by TGF-β. (A–C) Motif analysis (based on differential ATAC-peaks) of WT-TGFb vs. WT-SD Hyper ( p < 0.0001 and enrichment >2) (A) , and KO-TGFb vs. WT-TGFb Hypo ( p < 0.0001 and enrichment >2) (B) and KO-SD vs. WT-SD hypo ( p < 0.0001 and enrichment >2) (C) . (D) Proposed mechanisms of gene regulation by lncMGC mediated by SMARCA5. The promoters of TGF-β-regulated genes are open through Smad and ZF ARID sites even before TGF-β treatment. TGF-β-regulated genes are upregulated by the recruitment of Smad at open chromatin regions. (E) In the absence of lncMGC, the promoters of TGF-β-regulated genes are closed even after TGF-β treatment, and their expression is not increased even with promoter acetylation or TGF-β treatment. Interaction of lncMGC RNA with nucleosome remodelers can enhance the gene expression through the opening of chromatin at the promoter regions of TGF-β-regulated genes.

Article Snippet: The cross-linked chromatin was sheared and immunoprecipitated with antibodies against H3K27Ac (Abcam, ab4729) and SMARCA5 (Boster Biol.

Techniques: Expressing, Gene Expression

Journal: American Journal of Translational Research

Article Title: Endophilin A2 attenuates cardiac hypertrophy induced by isoproterenol through the activation of autophagy

doi:

Figure Lengend Snippet: EndoA2 attenuated hypertrophic responses induced by ISO in neonatal rat cardiomyocytes (NRCMs). A, B. NRCMs were transfected with Ad-EndoA2 or siRNA followed by incubation with ISO (1 μM for 24 h). Morphological changes were observed by staining with α-actinin. ISO increased cell surface area, whereas EndoA2 overexpression decreased the ISO-induced cell surface area, and EndoA2 siRNA knockdown further enhanced the effects of ISO on the cell surface area. n=6, *P<0.05 vs. control group, #P<0.05 vs. ISO group. C. The mRNA expression levels were determined by qPCR. Statistical analyses showed that ISO increased the mRNA expression levels of ANF, BNP and β-MHC, whereas EndoA2 overexpression significantly decreased the ISO-induced increases in ANF, BNP and β-MHC mRNA expression levels in NRCMs, EndoA2 siRNA knockdown further enhanced the effects of ISO on the mRNA expression levels of ANF, BNP and β-MHC. n=6, *P<0.05 vs. control group, #P<0.05 vs. ISO group.

Article Snippet: Following overnight incubation with α-actinin (Boster Biological Technology, Wuhan, CN), the cells were incubated with the corresponding secondary antibodies (Cell Signaling, Boston, MA) and the nuclei were stained with DAPI solution (Beyotime, Nanjing, CN).

Techniques: Transfection, Incubation, Staining, Over Expression, Expressing