Journal: Scientific Reports

Article Title: Abnormal Hypermethylation of the VDAC2 Promoter is a Potential Cause of Idiopathic Asthenospermia in Men

doi: 10.1038/srep37836

Figure Lengend Snippet: ( A ) psi_CHECK-2. ( B ) M: 3000 bp DNA Marker, 1–4 represent the brands of psi_CHECK-2-VDAC2 and replied with 5–8. The well of 1, 2, 3, 5, 6 and 7 were the fractional products of the VDAC2 promoter region; the well of 4 and 8 were the integrated products. ( C ) Partial sequence of the product of the recombined VDAC2 promoter region.

Article Snippet: A psi_CHECK-2 vector was purchased from Novobio, Shanghai ( ).

Techniques: Marker, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Proper Balance of Small GTPase rab10 Is Critical for PGC Migration in Zebrafish

doi: 10.3390/ijms222111962

Figure Lengend Snippet: Heatmap of representative DEGs related to cell migration and verification of the expression patterns by qPCR analysis. ( A ) Two-dimensional hierarchical cluster of DEGs associated with cell migration. ( B – M ) Validation of the transcriptome by qRT-PCR analysis of representative genes including ( B ) anxa1c , ( C ) cldne , ( D ) mcf2a , ( E ) megf6a , ( F ) myl7 , ( G ) rab10 , ( H ) rab36 , ( I ) hmgxb4a , ( J ) synpo2lb , ( K ) prdm12b , ( L ) dnd and ( M ) gra . The FPKM values (left) and the relative expression levels of qPCR (right) are shown. ( B – G ) The results were representative of more than three independent experiments in triplicate. Three reference genes ef1a , β-actin and rpl13a were used in every PCR examination to normalize gene expression levels with 2 -∆∆Ct method. The statistics were calculated and analyzed by Student’s t -test, * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: At 70–80% confluence, HEK293T cells seeded in 24-well plates were transiently co-transfected 10 ng psiCHECK2-3’UTR-prdm12b-WT ( psiCHECK2-3’UTR-rab10-WT ) or psiCHECK2-3’UTR-prdm12b-seed-Mut ( psiCHECK2-3’UTR-rab10-seed-Mut ) plasmids and 100nM control or miR-202-5p mimics with Lipofectamine 8000 (Beyotime, Shanghai, China).

Techniques: Migration, Expressing, Biomarker Discovery, Quantitative RT-PCR, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Proper Balance of Small GTPase rab10 Is Critical for PGC Migration in Zebrafish

doi: 10.3390/ijms222111962

Figure Lengend Snippet: miR-202-5p directly regulates prdm12b and rab10 . ( A , B ) The sequence information of the 3′utr of prdm12b and rab10 . The canonical miR-202-5p binding sites and its mutant were shown in red and blue, respectively. ( C ) Relative luciferase activity in HEK293T cells transfected with psiCHECK2- 3′utr wt ( prdm12b or rab10 ) or psiCHECK2- 3′utr mut ( prdm12b or rab10 ) in the presence of control mimics (con mimics) or miR-202-5p mimics. ( D ) qRT-PCR analysis of prdm12b and rab10 mRNA in 24hpf embryos injected with control or miR-202-5p mimics. ( E ) The representative images of embryos co-injected with mRNAs of rfp-prdm12b 3′utr wt , rfp-prdm12b 3′utr mut , rfp-rab10 3′utr wt or rfp-rab10 3′utr mut in the presence of control or miR-202-5p mimics. All embryos were co-injected with pEGFP-N3 vector as a control. ( F ) The relative signal ratio of RFP/GFP in the experiment presented in ( E ). ( C , D , F ) The results were representative of more than three independent experiments in triplicate. β-actin was used as an internal control to normalize gene expression levels with 2 -∆∆Ct method * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: At 70–80% confluence, HEK293T cells seeded in 24-well plates were transiently co-transfected 10 ng psiCHECK2-3’UTR-prdm12b-WT ( psiCHECK2-3’UTR-rab10-WT ) or psiCHECK2-3’UTR-prdm12b-seed-Mut ( psiCHECK2-3’UTR-rab10-seed-Mut ) plasmids and 100nM control or miR-202-5p mimics with Lipofectamine 8000 (Beyotime, Shanghai, China).

Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Transfection, Control, Quantitative RT-PCR, Injection, Plasmid Preparation, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Proper Balance of Small GTPase rab10 Is Critical for PGC Migration in Zebrafish

doi: 10.3390/ijms222111962

Figure Lengend Snippet: Sequences of primer used in this paper.

Article Snippet: At 70–80% confluence, HEK293T cells seeded in 24-well plates were transiently co-transfected 10 ng psiCHECK2-3’UTR-prdm12b-WT ( psiCHECK2-3’UTR-rab10-WT ) or psiCHECK2-3’UTR-prdm12b-seed-Mut ( psiCHECK2-3’UTR-rab10-seed-Mut ) plasmids and 100nM control or miR-202-5p mimics with Lipofectamine 8000 (Beyotime, Shanghai, China).

Techniques: Sequencing