Journal: PLoS Pathogens

Article Title: Mgat1 -dependent N-glycosylation of Membrane Components Primes Drosophila melanogaster Blood Cells for the Cellular Encapsulation Response

doi: 10.1371/journal.ppat.1002819

Figure Lengend Snippet: Paired brightfield and FITC images of hemocytes stained with FITC-WGA at the indicated time after wasp attack reveals that WGA staining of hemocytes is dynamic following wasp attack. (A,B) Plasmatocytes from unattacked w 1118 larvae, age matched with the 24–48 hour post attack (PA) time point, did not stain with WGA. (C,D) At 0–24 hours PA, lamellocytes from L. clavipes attacked w 1118 larvae began to show WGA staining. (E–H) A strong, speckled, WGA staining pattern was seen in lamellocytes (E,F), and to a lesser extent, plasmatocytes (G,H) from attacked w 1118 larvae at 24–48 hours PA. (I,J) By 48–72 hours PA, lamellocytes from attacked w 1118 larvae no longer stained with WGA. (K,L) Lamellocytes from attacked Mgat1 1 mutant larvae did not stain with WGA at 24–48 hours PA, demonstrating that the WGA signal is dependent on N-glycosylation pathway function. Scale bar in all panels indicate 10 µm.

Article Snippet: Hemocytes were then stained with 100 µg/ml FITC labeled WGA from Vector Laboratories (FL-1021) for 3 minutes and washed three times with Drosophila Ringer's solution as described in .

Techniques: Staining, Mutagenesis

Journal: PLoS Pathogens

Article Title: Mgat1 -dependent N-glycosylation of Membrane Components Primes Drosophila melanogaster Blood Cells for the Cellular Encapsulation Response

doi: 10.1371/journal.ppat.1002819

Figure Lengend Snippet: (A) Proportion of wasp eggs encapsulated in the indicated genotypes. Mgat1 appears haploinsufficient for encapsulation of both L. clavipes (left) and Aphaereta sp. (right) eggs. (B,C) Paired brightfield and FITC images of hemocytes stained with FITC-WGA. Lamellocytes from L. clavipes attacked Mgat1 1 /+ larvae show intermediate levels of WGA staining at 24–48 hours PA. Scale bar indicates 10 µm. (D) Proportion of wasp exposed larvae of the indicated genotypes that eclosed as adult flies. The decreased encapsulation rate of Mgat1 1 /+ larvae correlated with a decrease in fly eclosion. Error bars indicate standard error of the mean; * indicates p<0.01 relative to control.

Article Snippet: Hemocytes were then stained with 100 µg/ml FITC labeled WGA from Vector Laboratories (FL-1021) for 3 minutes and washed three times with Drosophila Ringer's solution as described in .

Techniques: Staining

Journal: PLoS Pathogens

Article Title: Mgat1 -dependent N-glycosylation of Membrane Components Primes Drosophila melanogaster Blood Cells for the Cellular Encapsulation Response

doi: 10.1371/journal.ppat.1002819

Figure Lengend Snippet: Paired brightfield and FITC images of hemocytes stained with FITC-WGA 24–48 hours following attack by the indicated wasp. (A–D) Lamellocytes from He-Gal4;w 1118 larvae were WGA+ following L. clavipes attack (A,B), but were WGA− following L. victoriae attack (C,D). (E,F) The WGA+ staining was restored to L. victoriae attacked larvae by hemocyte specific expression of Mgat1 in He-Gal4;UAS-Mgat1 larvae. Scale bars indicate 10 µm.

Article Snippet: Hemocytes were then stained with 100 µg/ml FITC labeled WGA from Vector Laboratories (FL-1021) for 3 minutes and washed three times with Drosophila Ringer's solution as described in .

Techniques: Staining, Expressing