Journal: PLoS ONE

Article Title: Identification and Characterization of Proteins Involved in Nuclear Organization Using Drosophila GFP Protein Trap Lines

doi: 10.1371/journal.pone.0053091

Figure Lengend Snippet: Panels (A–C) depict diploid cells from third instar larvae imaginal tissue of the CG11138 protein-trap allele labeled with DAPI (A), α-GFP (B) and merged (C) indicating that CG11138-GFP is localized in punctate bodies mainly around the nuclear periphery. Panels (D–G) show diploid cells from OR third instar larvae imaginal tissue labeled with DAPI (D), α-CG11138 (E), α-Lamin C to define the nuclear periphery (F) and the merged image (G). Panels (H–K) show DAPI (H), α-CG11138 (I), α-LaminC (J) and merged (K) images of diploid cells from third instar larvae imaginal tissue of lamC SZ18 /lamC SZ18 mutant flies. Loss of Lamin C disrupts the localization of CG11138.

Article Snippet: The following primary antibodies were used: rabbit α-GFP (Molecular Probes) 1∶400, rat α-Stwl (gift from Dr. Dennis McKearin) 1∶20, guinea pig α-Stwl 1∶800, rabbit α-CG11138 1∶1000, rabbit α-Df31 1∶400, rabbit α-dNlp (gift from Dr. James Kadonaga) 1∶200, mouse α-Lamin Dm0 (Developmental Studies Hybridoma Bank) 1∶200, mouse α-Lamin C (Developmental Studies Hybridoma Bank) 1∶200, rabbit α-CP190 1∶4000, rat α-Vasa (Developmental Studies Hybridoma Bank) 1∶200, rat α-Mod(mdg4)2.2 1∶1000, rabbit α-Su(Hw) 1∶2000, mouse α-MSL1 (gift from Dr. John Lucchesi), rabbit α-histone H3S10 ph (Millipore), rabbit α- phospho-Pol IIser5 (H14, Covance) and guinea pig α-CTCF 1∶2000.

Techniques: Labeling, Mutagenesis

Journal: PLoS ONE

Article Title: Identification and Characterization of Proteins Involved in Nuclear Organization Using Drosophila GFP Protein Trap Lines

doi: 10.1371/journal.pone.0053091

Figure Lengend Snippet: (A–C) dNLP-GFP flourescence in diploid cells from third instar larvae imaginal tissue from a dNLP protein-trap allele; DAPI (A), dNLP-GFP (B) and merged (C). Panels (D–G) depict dNLP localization in both somatic and germline cell nuclei including the oocyte nucleus (arrowhead in F and G); (D) DAPI, (E) α- Lamin Dm0, (F) α-dNLP and (G) merged. Panels H and I show dNLP-GFP fluorescence in an egg from a dNLP protein-trap allele (H) as compared to a wild type egg (I) indicating that dNLP-GFP is being dumped into the developing egg of the protein-trap allele. Panels J-N show Kc cells labeled with DAPI- blue, α-dNLP- red and α-H3S10ph-green in interphase (J), prophase (K), metaphase (L), anaphase (M) and telophase (N); the results suggest that dNLP is not associated with condensed chromosomes during metaphase. Panels (O–V). (O–R) show polytene chromosomes from wild type third instar larvae prior to heat shock while (S–V) show chromosomes from larvae subjected to a 20 min heat shock at 37°C. dNLP is broadly present in interbands and frequently colocalizes with RNA Pol II phosphorylated at Ser5 on the non-heats hocked polytene chromosomes. dNLP seems to become more diffuse and dissociate from the DNA following heat shock. The distribution of dNLP is particularly weak at the heat shock puffs, which are the only sites of transcription following temperature elevation (S–V). (O and S)-DAPI, (P and T)-α-dNLP, (Q and U)-α-PolII ser5 and (R and V)-merged.

Article Snippet: The following primary antibodies were used: rabbit α-GFP (Molecular Probes) 1∶400, rat α-Stwl (gift from Dr. Dennis McKearin) 1∶20, guinea pig α-Stwl 1∶800, rabbit α-CG11138 1∶1000, rabbit α-Df31 1∶400, rabbit α-dNlp (gift from Dr. James Kadonaga) 1∶200, mouse α-Lamin Dm0 (Developmental Studies Hybridoma Bank) 1∶200, mouse α-Lamin C (Developmental Studies Hybridoma Bank) 1∶200, rabbit α-CP190 1∶4000, rat α-Vasa (Developmental Studies Hybridoma Bank) 1∶200, rat α-Mod(mdg4)2.2 1∶1000, rabbit α-Su(Hw) 1∶2000, mouse α-MSL1 (gift from Dr. John Lucchesi), rabbit α-histone H3S10 ph (Millipore), rabbit α- phospho-Pol IIser5 (H14, Covance) and guinea pig α-CTCF 1∶2000.

Techniques: Fluorescence, Labeling

Journal: PLoS ONE

Article Title: Identification and Characterization of Proteins Involved in Nuclear Organization Using Drosophila GFP Protein Trap Lines

doi: 10.1371/journal.pone.0053091

Figure Lengend Snippet: Panels (A–F) show GFP fluorescence for Stwl-GFP in polytene chromosomes (A–C) and in diploid cells (D–F). (A and D) show DNA labeled in blue by DAPI. Stwl-GFP is shown in green in panels (B and E) while panels (C and F) show the merged images. Stwl localization appears as dots around the periphery of diploid cells; DAPI (G), α-Stwl (H), α-Lamin Dm0 (I) and merged (J). Panels (K–N) show polytene chromosomes labeled with Stwl and CP190 antibodies indicating little overlap between the two proteins, DAPI (K), α-Stwl (L), α-CP190 (M) and merged (N).

Article Snippet: The following primary antibodies were used: rabbit α-GFP (Molecular Probes) 1∶400, rat α-Stwl (gift from Dr. Dennis McKearin) 1∶20, guinea pig α-Stwl 1∶800, rabbit α-CG11138 1∶1000, rabbit α-Df31 1∶400, rabbit α-dNlp (gift from Dr. James Kadonaga) 1∶200, mouse α-Lamin Dm0 (Developmental Studies Hybridoma Bank) 1∶200, mouse α-Lamin C (Developmental Studies Hybridoma Bank) 1∶200, rabbit α-CP190 1∶4000, rat α-Vasa (Developmental Studies Hybridoma Bank) 1∶200, rat α-Mod(mdg4)2.2 1∶1000, rabbit α-Su(Hw) 1∶2000, mouse α-MSL1 (gift from Dr. John Lucchesi), rabbit α-histone H3S10 ph (Millipore), rabbit α- phospho-Pol IIser5 (H14, Covance) and guinea pig α-CTCF 1∶2000.

Techniques: Fluorescence, Labeling