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Image Search Results
Journal: The Journal of Reproduction and Development
Article Title: C-X-C chemokine receptor type 4 (CXCR4) is a key receptor for chicken primordial germ cell migration
doi: 10.1262/jrd.2017-067
Figure Lengend Snippet: Experimental design of chicken C-X-C chemokine receptor type 4 ( CXCR4 ) gene targeting. (A) Reverse transcription polymerase chain reaction (RT-PCR) analysis of CXCR4 transcript in different chicken cell types (primordial germ cells [PGCs], DT40, and DF1). PGCs and DT40 cells strongly express CXCR4, but DF1 cells do not. (B) Schematic of CRISPR targeting of the chicken CXCR4 gene on chromosome 7 and genomic mutation analysis. Sequences of the guide RNA target site are located in the second exon (yellow box). TGG in red is a protospacer adjacent motif (PAM). Lowercase and capital letters are intron and exon sequences, respectively. Green boxes denote primer annealing sites. (C) Mutated target DNA sequences of chicken CXCR4 KO PGC line #3 induced by CRISPR/Cas9. Dashes denote deleted nucleotides. Mutant genotypes show a frameshift in translated proteins that induces amino acid changes and a stop codon (asterisk). (D) Genotyping analysis of CXCR4 RT-PCR product amplified from CXCR4 KO PGC line #3.
Article Snippet: To generate GFP- or DsRed-expressing PGCs, DF1, and
Techniques: Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, CRISPR, Mutagenesis, Amplification
Journal: The Journal of Reproduction and Development
Article Title: C-X-C chemokine receptor type 4 (CXCR4) is a key receptor for chicken primordial germ cell migration
doi: 10.1262/jrd.2017-067
Figure Lengend Snippet: Establishment of stably-transfected cells. (A) Schematic diagrams of transgene expression vectors for enhanced green fluorescent protein (eGFP), Discosoma red fluorescent protein (DsRed), and chicken CXCR4 controlled by the cytomegalovirus (CMV) promoter. Constructs were inserted between 5’-terminal repeat (5’-TR) and 3’-TR piggy Bac transposon elements. The neomycin-resistant (Neo R ) gene was used as a selection marker. (B) RT-PCR analysis after transfection and selection of piggy Bac CMV-chicken CXCR4-overexpressing (OE) DF1 cells. (C) Fluorescent images of GFP or DsRed-expressing wild type and GFP-expressing CXCR4 KO PGCs. All PGC sublines strongly expressed the fluorescent reporters (magnification, 100 ×). (D) GFP-expressing DF1 and DT40 cells after transfection and selection of the piggy Bac CMV-eGFP expression vector (magnification, 100 ×).
Article Snippet: To generate GFP- or DsRed-expressing PGCs, DF1, and
Techniques: Stable Transfection, Transfection, Expressing, Construct, Selection, Marker, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation
Journal: The Journal of Reproduction and Development
Article Title: C-X-C chemokine receptor type 4 (CXCR4) is a key receptor for chicken primordial germ cell migration
doi: 10.1262/jrd.2017-067
Figure Lengend Snippet: In vivo migration assay of GFP-expressing DT40 and CXCR4 OE DF1 cells. GFP-expressing DT40 or DF1 cells carrying the CMV-controlled chicken CXCR4-overexpressing transgene were transplanted into the bloodstream of recipient embryos. Embryonic gonads were dissected on Day 6 (A) and Day 10 (B). GFP-expressing cells were examined using a fluorescent microscope.
Article Snippet: To generate GFP- or DsRed-expressing PGCs, DF1, and
Techniques: In Vivo, Migration, Expressing, Microscopy