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Image Search Results
Journal: The Journal of Biological Chemistry
Article Title: Inhibition of Nucleotide Sugar Transport in Trypanosoma brucei Alters Surface Glycosylation
doi: 10.1074/jbc.M113.453597
Figure Lengend Snippet: TbNST4 is localized at the Golgi apparatus. A, genomic PCR analysis of in situ C-terminal HA-tagged tbnst4 in T. brucei BSF. Genomic DNA was isolated from untagged (lane 1) and tagged (lane 2) TbNST4 cells. In situ tagging locus was PCR amplified with a forward primer complementary to TbNST4 coding sequence and a reverse primer complementary to the 3′ UTR (outside the recombination sequence). The expected genomic PCR products are indicated. B, Western blot analysis of the in situ tagged TbNST4HA protein expression. Cell lysates from untagged (lane 1) and tagged TbNST4HA (lane 2) cells were subjected to SDS-PAGE and Western blotting with anti-HA antibody detection. The position of the expressed TbNST4 is indicated. C, immunofluorescence analysis of TbNST4HA localization. BSF cells carrying the tagged TbNST4 were stained with anti-HA for TbNST4 (green), anti-Ty for TbGT15 (red, upper panel), or anti-p67 (red, lower panel) antibodies. Cell images were captured with Nikon wide-field epifluorescence microscope. Blue, DAPI staining for nuclei and kinetoplasts.
Article Snippet: Cell images were captured with a
Techniques: In Situ, Isolation, Amplification, Sequencing, Western Blot, Expressing, SDS Page, Immunofluorescence, Staining, Microscopy