Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: The abnormal m6A modification and protein levels of GLIS1 in human aged kidney tissues. A Total m6A RNA levels in the H/aged ( n = 10) and H/control group ( n = 10) detected by m6A methylation quantification kit; B differential methylated mRNA expressions between H/aged (A) and H/control (Y) group by using m6A-mRNA epitranscriptomic microarray analysis ( n = 3); C analysis of significant changes in methylated mRNA expression levels between the H/aged and H/control group; D the Gene Ontology (GO) biological process enrichment analysis pathways of predicted mRNA targets; E enriched m6A modification of GLIS1 in the H/aged and H/control group by MeRIP assay ( n = 4); F , G protein level of GLIS1 in the H/aged and H/control group by western blot, and its semi-quantitative analyses ( n = 6). The data are expressed as the mean ± SD of three independent experiments. *** P < .001 indicates a significant difference versus the H/control group by Student’s t -test

Article Snippet: Human m6A epitranscriptomic microarray and mRNA microarray analyses were performed based on Arraystar’s standard protocols.

Techniques: Modification, Control, Methylation, Microarray, Expressing, Western Blot

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: METTL3 directly interacted with GLIS1 in kidney aging. A The mRNA level of METTL3 in the vector and siMETTL3 group in HK-2 cells by RT-qPCR ( n = 3); B , C protein level of GLIS1 by western blot in the vector and siMETTL3 group in HK-2 cells, and its semi-quantitative analysis ( n = 3); D protein levels of METTL3, METTL14, and WTAP by western blot in the 24-month-old and 6-month-old group ( n = 6); E , F the expression of METTL3 in the 24-month-old and 6-month-old group by IHC assay, and its semi-quantitative analysis ( n = 6), scale bar = 50 μm; G immunostaining for METTL3 (green), with DAPI (blue) counterstaining by IF staining in the 24-month-old and 6-month-old group ( n = 6), scale bar = 50 μm; H immunostaining for METTL3 (green), GLIS1 (red), with DAPI (blue) counterstaining by IF staining in HK-2 cells ( n = 3), scale bar = 50 μm; I , J METTL3 RIP and RT-PCR confirmed the interaction between METTL3 and GLIS1 mRNA, and its semi-quantitative analysis ( n = 3); K the lower m6A level of GLIS1 in siMETTL3 group compared with the vector group in HK-2 cells by using MeRIP-qPCR ( n = 3); L the mRNA level of GLIS1 in the vector and siMETTL3 group in HK-2 cells by RT-qPCR ( n = 3); M mutations at the two putative m6A sites in GLIS1 (A to G); N m6A level of GLIS1 in HK-2 cells with co-expression of siMETTL3 and GLIS1-WT/Muts by MeRIP-qPCR ( n = 3); O the mRNA level of GLIS1 in the vector and GLIS1-Mut3 group by RT-qPCR ( n = 3). The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the vector group ( A , C , K ) or 6-month-old group ( E ) or IgG group ( J ) by Student’s t -test; ## P < .01, ### P < .001 versus the vector-WT group, ++ P < .01, +++ P < .001 versus the siMETTL3-WT group, ** P < .01 versus vector group by two way-ANOVA ( N )

Article Snippet: Human m6A epitranscriptomic microarray and mRNA microarray analyses were performed based on Arraystar’s standard protocols.

Techniques: Plasmid Preparation, Quantitative RT-PCR, Western Blot, Expressing, Immunostaining, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: METTL3 ameliorated age-related renal fibrosis in accelerated aging mouse model. A , B Downregulated protein levels of METTL3, METTL14, and GLIS1 in an accelerated aging mouse model were reversed in the presence of AAV-METTL3, and the upregulated protein levels of P16 INK4A and FN by western blot in the accelerated aging mouse model were reduced by introducing AAV-METTL3 ( n = 6); C the enriched m6A modification of GLIS1 in the control, AAV-Vector and AAV-METTL3 group by MeRIP-qPCR assay ( n = 6); D GLIS1 mRNA levels in the control, AAV-Vector, and AAV-METTL3 group detected by RT-qPCR ( n = 5); E protein levels of GLIS1, P16 INK4A , and FN in the control, AAV-Vector and AAV-METTL3 group by Masson staining and IHC assay and their semi-quantitative analyses ( n = 6), scale bar = 50 μm; F immunostaining for PPARα (red) and CPT1A (green), HK2 (pink), and PDK1 (yellow), with DAPI (blue) counterstaining by IF staining in the control, AAV-Vector, and AAV-METTL3 group ( n = 6), scale bar = 50 μm. The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the AAV-vector group by one-way ANOVA

Article Snippet: Human m6A epitranscriptomic microarray and mRNA microarray analyses were performed based on Arraystar’s standard protocols.

Techniques: Western Blot, Modification, Control, Plasmid Preparation, Quantitative RT-PCR, Staining, Immunostaining

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: YTHDF1 identified with m6A-meditated GLIS1 mRNA and participated the translation process of GLIS1 protein. A The expression of YTHDF1 in the 24-month-old and 6-month-old group by IHC assay, and its semi-quantitative analysis ( n = 6), scale bar = 50 μm; B immunostaining for YTHDF1 (green), with DAPI (blue) counterstaining by IF staining in the 24-month-old and 6-month-old group ( n = 6), scale bar = 50 μm; C , D protein level of YTHDF1 in the 24-month-old and 6-month-old group by western blot, and its semi-quantitative analysis ( n = 6); E mRNA level of YTHDF1 by RT-qPCR in HK-2 cells within vector or siYTHDF1 ( n = 3); F protein level of GLIS1 in HK-2 cells within vector or siYTHDF1, and its semi-quantitative analysis ( n = 3); G double immunostaining for GLIS1 (red) and YTHDF1 (green) by IF staining in HK-2 cells ( n = 3), scale bar = 50 μm; H , I RIP and RT-PCR assays confirmed the interaction between YTHDF1 and GLIS1 mRNA, and its semi-quantitative analysis ( n = 3); J the expression of GLIS1 with RPL22-FLAG label by ribosomal immunoprecipitation in HK-2 cells ( n = 3); K the mRNA level of GLIS1 by RT-qPCR in HK-2 cells within vector or siYTHDF1 ( n = 3); L , M relative luciferase activity of the GLIS1-WT or GLIS1-Mut 3′UTR luciferase reporter in the vector and siMETTL3 group, n = 3. The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the 6-month-old group, or vector group, or IgG group by Student’s t -test

Article Snippet: Human m6A epitranscriptomic microarray and mRNA microarray analyses were performed based on Arraystar’s standard protocols.

Techniques: Expressing, Immunostaining, Staining, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Double Immunostaining, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation, Luciferase, Activity Assay

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A) Predicted AP-1 (yellow) and KLF (framed) binding sites on mouse Ppara promoter. (B–C) Cardiac mRNA levels of Klf isoforms (B) and protein levels of KLF5 and β-actin (C) in 10–12-weeks old C57BL/6 mice treated with 5 mg/kg LPS or saline (CTRL) (n=4–5; *P<0.05; **P<0.01; ***P<0.001 vs CTRL). (D–E) Ppara, Klf5 and Klf6 mRNA levels in HL-1 cells (D) treated with 1µg/ml LPS or saline (CTRL) for 9h (n=6; *p<0.05 vs. CTRL) or in aMHC-Pparg mice (E) treated with 5mg/kg LPS or saline (CTRL) for 8–10h (n=5; *p<0.05; **p<0.01 vs. CTRL).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques: Binding Assay, Saline

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A–D) Ppara and Klf5 mRNA (A, C) and protein (B, D) levels in HL-1 cells treated with Ad-cJunAsp (A, B) or Ad-KLF5 (C, D); (n=6; *p<0.05; **p<0.01; ***p<0.001 vs CTRL). (E–I) Enrichment of −792/−772 bp region (E, F) or −719/−698 bp region (G, H) of mouse Ppara promoter with c-Jun (E, G) or KLF5 (F, H) of chromatin samples from HL-1 cells treated with Ad-GFP (CTRL) and either Ad-cJunAsp (E, G) or Ad-KLF5 (F, H); **p<0.01 vs CTRL. (I) Enrichment of −792/−772 bp region of mouse Ppara promoter with c-Jun or KLF5 of chromatin samples from HL-1 cells treated with 1 µg/ml LPS or saline (CTRL); *p<0.05 vs CTRL. Data for all bar graphs are represented as means ± SEM (statistical analysis: t-test).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques: Saline

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A, B) Klf5 mRNA in the heart, skeletal muscle, intestine, kidney, white adipose tissue, brain (A) and primary cardiac myocytes (B) of aMHC-Klf5−/− mice (n=3; *p<0.05 vs floxed). (C) Hierarchical clustering for differentially expressed mRNAs detected by whole genome microarray analysis of cardiac mRNA obtained from aMHC-Klf5−/− mice and control floxed mice. Red color indicates high relative expression and blue color indicates low relative expression. (D–G) Gene ontology analysis for classification of the downregulated (D) or upregulated (E) genes based on the metabolic process that they are associated with and pathway analysis for downregulated (F) and upregulated (G) genes detected with whole genome microarray analysis of cardiac mRNA obtained from aMHC-Klf5−/− mice and control floxed mice. Data for all bar graphs are represented as means ± SEM (statistical analysis: t-test).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques: Microarray, Control, Expressing

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A) Ingenuity pathway analysis of genes regulated over 2-fold in the aMHC-Klf5−/− mouse array that are related to FA metabolism. (B) Cardiac Klf5 and Ppara mRNA levels of 10- to 12-week-old aMHC-Klf5−/− male and female mice (n=5; **p<0.01; ***p<0.001 vs same gender floxed mice). (C) Cardiac PPARα and β-actin protein levels of 10- to 12-week-old floxed and aMHC-Klf5−/− male mice. (D–F) Cardiac mRNA levels for FA oxidation- (Ppargc-1a, Ppargc-1β, Pparg, Ppard, Acox and Cpt1b) (D), lipid uptake- (Cd36, Lpl and Angptl4) (E) and lipid storage-related genes (Dgat1, Dgat2, Plin2, Plin5) (F) (n=5; *p<0.05, **p<0.01, ***p<0.001 vs same gender floxed mice). (G) Cardiac PGC-1, CPT-1, DGAT-1, ATGL, phosphorylated AMPK, total AMPK, and GAPDH protein levels of 10- to 12-week-old floxed and aMHC-Klf5−/− male mice. (H, I) [14C]-Palmitic acid (H) and [14C]-Glucose (I) oxidation levels in cardiac muscle of 10- to 12-week-old floxed and aMHC-Klf5−/− male mice (n=4–5; *p<0.05; **p<0.01 vs floxed mice). Data for all bar graphs are represented as means ± SEM (statistical analysis: t-test).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques:

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A–F) Fractional shortening (A, D), left ventricular internal dimension during diastole (B, E), left ventricular internal dimension during systole (C, F), in 2–3 months old (A–C) and 6 months old (D–F) αMHC-Klf5−/− and floxed (WT) mice. (G–M) Photographs of echocardiograms (G), fractional shortening (H), left ventricular internal dimension during diastole (I), left ventricular internal dimension during systole (J) left ventricular posterior wall during diastole (K) left ventricular posterior wall during systole (L), and heart weight/tibia length ratio (M) in 8–12 months old αMHC-Klf5−/− and floxed (WT) mice (n=7–8; *p<0.05). (N, O) Cardiac mRNA levels for Bnp, Anf, αMHC and βMHC genes in 2–3 months old (N) and 11–12 months old (O) male floxed and αMHC-Klf5−/− mice (F) (n=5; *p<0.05, **p<0.01 vs floxed mice).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques:

Journal: Circulation research

Article Title: Cardiac Myocyte KLF5 Regulates Ppara Expression and Cardiac Function

doi: 10.1161/CIRCRESAHA.115.306383

Figure Lengend Snippet: (A) Ingenuity pathway analysis of cardiac genes regulated over 2-fold in the aMHC-Klf5−/− mouse array that have direct or indirect association with insulin signaling and glucose metabolism proteins. Highlighted with bold fonts within the diagram are proteins that modulate insulin signaling. (B) Fractional shortening of C57BL/6 mice 6 weeks post-STZ or saline (CTRL) administration (n=5; *p<0.05 vs CTRL). (C) Western blot analysis for cardiac KLF5 and β-actin protein levels in C57BL/6 mice 6 weeks post-STZ administration (n=3; ***p<0.001 vs CTRL). (D) Cardiac Klf5 and Ppara mRNA levels in floxed and aMHC-Klf5−/− mice 6 weeks post-STZ administration (n=5; *p<0.05, **p<0.01 vs CTRL). (E) Cardiac Klf5 and Ppara mRNA levels in 12 weeks old ob/ob mice compared with wild type C57BL/6 mice (n=4–5, *p<0.05, ***p<0.001 vs wt). (F–I) Plasma glucose levels (F, G) and cardiac Klf5 and Ppara mRNA levels (H, I) in wild type mice treated with STZ (6 weeks prior to glucose measurement), dapagliflozin (F, H), antisense oligonucleotides against SGLT2 (SGLT2-ASO) (G, I) and combination of either STZ with dapagliflozin (F, H) or STZ with SGLT2-ASO (G, I) (n=5, **p<0.01, ***p<0.001 vs CTRL).

Article Snippet: The microarray analysis for cardiac mRNA obtained from α MHC-Klf5 −/− mice was performed by Arraystar (data deposited in {"type":"entrez-geo","attrs":{"text":"GSE63839","term_id":"63839"}} GSE63839 ).

Techniques: Saline, Western Blot, Clinical Proteomics

Journal: Cancer Management and Research

Article Title: LRH-1 drives hepatocellular carcinoma partially through induction of c-myc and cyclin E1, and suppression of p21

doi: 10.2147/CMAR.S162887

Figure Lengend Snippet: Suppression of LRH-1 in HepG2 prevents its growth and colony formation capacity in vitro. Notes: ( A ) Schematic diagram of the TALENs targeting the second exon of LRH-1 and Western blotting (WB) validation of HepG2 LRH-1/− cell clones. ( B ) Proliferation assay by MTT. Two HepG2 LRH-1/− cell clones proliferated at slower rates than the parental HepG2 cells. ( C ) Colony formation assay. Results showed that HepG2 LRH-1/− cells formed smaller and less colonies than parental HepG2 cells. ** P <0.01 vs parental HepG2 cells. Abbreviation: TALENs, transcription activator-like effector nucleases.

Article Snippet: HepG2 mRNA microarray was performed with Arraystar Human LncRNA Microarray v4.0 according to the manufacturer’s protocol.

Techniques: In Vitro, TALENs, Western Blot, Biomarker Discovery, Clone Assay, Proliferation Assay, Colony Assay

Journal: Cancer Management and Research

Article Title: LRH-1 drives hepatocellular carcinoma partially through induction of c-myc and cyclin E1, and suppression of p21

doi: 10.2147/CMAR.S162887

Figure Lengend Snippet: Suppression of LRH-1 in HepG2 attenuates its growth in vivo. Notes: ( A and B ) Growth curve shows the growth patterns of tumors formed by HepG2 LRH-1/− cell clones and parental HepG2 cells, HepG2 cells formed xenograft tumors with larger sizes and at faster rates as compared to HepG2 LRH-1/− #1 cells, while HepG2 LRH-1/− #2 lost the in vivo tumorigenic capacity in all nude mice (NT, no tumor). ( C ) Ki67 immunohistochemistry. Our results suggested that significantly lower and less Ki67 immunoreactivity was seen in xenograft tumors formed from HepG2 LRH-1/− than parental HepG2 cells. Scale bar =100 μm.** P <0.01 vs parental HepG2 cells.

Article Snippet: HepG2 mRNA microarray was performed with Arraystar Human LncRNA Microarray v4.0 according to the manufacturer’s protocol.

Techniques: In Vivo, Clone Assay, Immunohistochemistry

Journal: Cancer Management and Research

Article Title: LRH-1 drives hepatocellular carcinoma partially through induction of c-myc and cyclin E1, and suppression of p21

doi: 10.2147/CMAR.S162887

Figure Lengend Snippet: Gene expression microarray analysis for the identification of LRH-1-regulated genes in HepG2 cells. Notes: ( A ) Gene ontology (GO) analysis of genes that are LRH-1-regulated in HepG2 and in HepG2 LRH-1/− cells. ( B ) KEGG pathway enrichment analysis for LRH-1-regulated genes in HepG2 cells are shown. ( C ) Several cell cycle-related genes that altered after suppression of LRH-1 were listed. Abbreviations: BP, biological process; DE, differential expression; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: HepG2 mRNA microarray was performed with Arraystar Human LncRNA Microarray v4.0 according to the manufacturer’s protocol.

Techniques: Gene Expression, Microarray, Quantitative Proteomics

Journal: Cancer Management and Research

Article Title: LRH-1 drives hepatocellular carcinoma partially through induction of c-myc and cyclin E1, and suppression of p21

doi: 10.2147/CMAR.S162887

Figure Lengend Snippet: LRH-1 promotes the development and progression of HCC through induction of c-myc and cyclin E1, and suppression of p21. Notes: ( A ) qRT-PCR analysis of c-myc, cyclin E1, and p21 genes in HepG2 and HuH-7 cells with ectopic LRH-1 expression. Our data indicated that ectopic expression of LRH-1 dramatically induced the mRNA and protein levels of c-myc and cyclin E1, and attenuated, in the meanwhile, the expression of p21. ( B ) Western blotting validation of c-myc, cyclin E1, and p21 in HepG2 and HuH-7 with ectopic LRH-1 expression. ( C ) Reporter assay showed that luciferase activity of c-myc and cyclin E1 promoter-luc reporter was significantly enhanced, while that of p21 was dramatically suppressed, in HepG2-LRH-1 cells. ( D ) MTT results indicated that the increased growth ability of HepG2-LRH-1 cells could be, at least partially, reversed by suppressing the expression of c-myc or restoring the expression of p21. ( E ) Schematic diagram showing the hypothesized pathways of LRH-1-mediated cell growth in HCC via its transcriptional repression of p21 and transactivation of c-myc and cyclin E1. * P <0.05; ** P <0.01; # not significant. Abbreviations: HCC, hepatocellular carcinoma; luc, luciferase.

Article Snippet: HepG2 mRNA microarray was performed with Arraystar Human LncRNA Microarray v4.0 according to the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Biomarker Discovery, Reporter Assay, Luciferase, Activity Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: The Expression Pattern of PDGFR-β in the Fibrotic Liver of MCDHF Mice and the Correlation between PDGFR-β and Fibrosis and Angiogenesis Markers (A) Representative H&E, Sirius Red, and oil red O stainings in the fibrotic liver of MCDHF mice. The mRNA expressions of fibrosis markers (B), angiogenesis markers (C), and PDGFR-β (D) were examined by qRT-PCR in the fibrotic liver of MCDHF mice. PDGFR-β protein expression was examined (E) and quantified (F) by western blot in the fibrotic liver. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: The Origin of PDGFR-β + Cells in MCDHF Fibrotic Mice and the Expression of PDGFR-β in TGF-β1-Treated BMSCs In Vivo (A) Representative images of immunofluorescence analysis to track PDGFR-β (green) expression in the fibrotic liver. Hollow arrows indicate PDGFR-β expression around the newly formed vessels, while solid arrows indicate PDGFR-β expression around the existing mature vessels. (B) Immunofluorescence staining for PDGFR-β (red) to track PDGFR-β + cells following 56 days of MCDHF diet. (C) The proportion of PDGFR-β + EGFP + cells accounting for total PDGFR-β + cells was measured by Image-Pro Plus software. (D) PDGFR-β mRNA expression was examined by qRT-PCR in BMSCs treated with the indicated concentrations of TGF-β1 for 24 h. (E) PDGFR-β mRNA expression in BMSCs treated with 10 ng/mL TGF-β1 at different times. (F and G) PDGFR-β protein expression was examined (F) and quantified (G) by western blot in TGF-β1-treated BMSCs. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Expressing, In Vivo, Immunofluorescence, Staining, Software, Quantitative RT-PCR, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: Downregulation of PDGFR-β Expression by miR-26b-5p in TGF-β1-Treated BMSCs (A) miR-26b-5p expression in the fibrotic liver induced by MCDHF. (B) The correlation between miR-26b-5p and PDGFR-β in liver tissue. Transfection efficiency of miR-26b-5p mimic (C) or inhibitor (F) in BMSCs is shown. PDGFR-β mRNA expression was examined by qRT-PCR with or without miR-26b-5p mimic (D) or inhibitor (G) transfection in BMSCs. Protein expression was examined by western blot with miR-26b-5p mimic (E) or inhibitor (H) transfection in BMSCs. Biotin-avidin pull-down assay (I) and luciferase reporter assay (J) demonstrated PDGFR-β was a target of miR-26b-5p. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control. #p < 0.05 versus TGF-β1 treated alone.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Avidin-Biotin Assay, Pull Down Assay, Luciferase, Reporter Assay, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: Regulation of lncMEG3 on PDGFR-β in TGF-β1-Treated BMSCs (A) lncMEG3 expression was examined by qRT-PCR in the fibrotic liver induced by MCDHF. (B) The correlation between lncMEG3 and PDGFR-β in liver tissue. Transfection efficiency of lncMEG3 overexpression plasmid (C) or siRNA (F) in BMSCs is shown. PDGFR-β mRNA expression was examined by qRT-PCR with lncMEG3 overexpression plasmid (D) or siRNA (G) in BMSCs. PDGFR-β protein expression was examined by western blot with lncMEG3 overexpression plasmid (E) or siRNA (H) in BMSCs. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control. #p < 0.05 versus TGF-β1 treated alone.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: miR-26b-5p Was Sponged by lncMEG3 in TGF-β1-Treated BMSCs (A) The correlation between lncMEG3 and miR-26b-5p in liver tissue. Biotin-avidin pull-down assay (B) and luciferase reporter assay (C) demonstrated that lncMEG3 was a target of miR-26b-5p. (D) PDGFR-β mRNA expression with or without lncMEG3 overexpression plasmids in miR-26b-5p mimic-treated BMSCs. (E) PDGFR-β protein expression with or without lncMEG3 overexpression plasmids in miR-26b-5p mimic-treated BMSCs. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control. #p < 0.05 versus miR-26b-5p mimic treated alone.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Avidin-Biotin Assay, Pull Down Assay, Luciferase, Reporter Assay, Expressing, Over Expression, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: Microarray Analysis Was Performed in TGF-β1-Treated BMSCs with or without miR-26b-5p Mimics BMSCs were treated with 10 ng/mL TGF-β1 for 24 h with or without miR-26b-5p mimics. Microarray analysis for mRNA was performed with RNA extracted from BMSCs. Hierarchical cluster analysis of significantly differentially expressed mRNA is shown as follows: bright green, underexpression; black, no change; bright red, overexpression. n = 3 per group. (A) Microarray heatmap comparing the signatures of PDGFR-β and angiogenesis marker and fibrosis marker genes in TGF-β1-treated BMSCs with or without miR-26b-5p mimics. KEGG and Reactome enrichment analyses (B) and GO enrichment analysis for biological process terms (C) are for the differentially expressed genes with miR-26b-5p mimics. n = 3 per group. (D) Predicted protein network visualization with STRING. The network view predicted the associations between proteins from the regulated genes involved in angiogenesis and extracellular matrix organization in TGF-β1-treated BMSCs. The network nodes were proteins. These proteins were clustered using k-means clustering algorithms.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Microarray, Over Expression, Marker

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA-26b-5p Inhibits Mouse Liver Fibrogenesis and Angiogenesis by Targeting PDGF Receptor-Beta

doi: 10.1016/j.omtn.2019.02.014

Figure Lengend Snippet: The Regulation of miR-26b-5p Agomir on PDGFR-β Expression, Liver Fibrosis, and Angiogenesis In Vivo (A) Transfection efficiency of 50 nM miR-26b-5p agomir (mimic in vivo ) in the fibrotic liver of MCDHF mice. (B) PDGFR-β mRNA levels in liver tissue were measured by qRT-PCR with or without miR-26b-5p agomir injection in MCDHF mice. (C) PDGFR-β protein levels in liver tissue were measured by western blot. (D) Representative images of immunofluorescence analysis to track PDGFR-β (red) expression in BMSCs (green) in the fibrotic liver. (E) The proportion of PDGFR-β + EGFP + cells of total PDGFR-β + cells was measured by Image-Pro Plus software. Hollow arrows indicate PDGFR-β − EGFP + cells, while solid arrows indicate PDGFR-β + EGFP + cells. DAPI was used to visualize nuclei (blue). The mRNA levels of angiogenesis markers (F) and fibrosis markers (G) in the fibrotic liver are shown. Data are presented as the mean ± SEM. n = 6 per group. *p < 0.05 versus control. #p < 0.05 versus MCDHF treated alone.

Article Snippet: KEGG, Reactome, and GO analyses were performed to explore the function and pathways based on differentially expressed genes. mRNA microarray experiments and technical assistance in bioinformatic analysis were handled by Capitalbio Technology (Beijing, China).

Techniques: Expressing, In Vivo, Transfection, Quantitative RT-PCR, Injection, Western Blot, Immunofluorescence, Software, Control

Journal: OncoTargets and therapy

Article Title: Long Noncoding RNA LINC01485 Promotes Tumor Growth and Migration via Inhibiting EGFR Ubiquitination and Activating EGFR/Akt Signaling in Gastric Cancer

doi: 10.2147/OTT.S257151

Figure Lengend Snippet: Microarray analysis of differentially expressed lncRNAs and mRNAs in gastric cancer. ( A ) Heat map and hierarchical clustering analyzed the distinguishable lncRNA ( A ) and mRNA ( B ) profiling in gastric cancer tumor tissues and paratumor counterparts. The relative expression from high to low level was indicated with red and blue color. ( C and D ) The expression levels of 6 upregulated candidate mRNAs and 8 upregulated novel candidate lncRNAs were validated in 20 paired gastric cancer tumor tissues and paratumor counterparts with RT-PCR. The data were presented as the mean ± SEM, * P < 0.05, Student’s t -test. ( E ) The specific gene-expression patterns were determined by weighted gene co-expression network analysis (WGCNA). ( F ) The correlation analysis of EGFR and 4 lncRNAs were examined in 20 gastric cancer tumor tissues with RT-PCR. Abbreviations: lncRNA, long non-coding RNA; RT-PCR, real-time polymerase chain reaction; SEM, standard error of the mean; EGFR, epithelial growth factor receptor.

Article Snippet: The lncRNA/mRNA crosslinked microarray was employed (Capitalbio, Beijing, China).

Techniques: Microarray, Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Real-time Polymerase Chain Reaction