1. Volume 4, Issue 1, Pages 129-135 (July 1999)
JIL-1 
Ye Jin, Yanming Wang, Diana L Walker, Hao Dong, Cherice Conley, Jørgen Johansen, Kristen M Johansen 
Molecular Cell 
Volume 4, Issue 1, Pages (July 1999)
DOI: /S (00)

2. Figure 1
The Predicted Protein Sequence and Phylogenetic Relationship of JIL-1
(A) The complete 3621 nt open reading frame derived from the JIL-1 cDNA sequence translates into a 1207–amino acid protein with a predicted molecular mass of 137 kDa. The shaded boxes in the sequence indicate the two domains with homology to serine/threonine kinase catalytic domains. The sequence also includes a putative bipartite nuclear localization signal (boxed region) and three underlined regions that show significantly high PEST sequence prediction scores (11.90, 12.98, and 11.58, respectively).
(B) Schematic diagrams of JIL-1, human MSK1, and Drosophila RSK drawn to scale to compare the domain organization of these tandem kinases.
Molecular Cell 1999 4, DOI: ( /S (00) )

3. Figure 2 JIL-1 Immunoblots and In Vitro JIL-1 Kinase Assays
(A) Immunoblots of SDS-PAGE-fractionated embryonic Drosophila proteins labeled with Odin and Hope antisera against JIL-1. Both antisera recognize a doublet or triplet of bands with relative molecular masses of 150–160 kDa (lanes 1 and 2).
(B) In vitro JIL-1 kinase assay. JIL-1 was immunoprecipitated from Schneider-2 cells using Odin antiserum (Odin ip). A portion was added to an in vitro kinase assay, fractionated by SDS-PAGE, stained, dried, and autoradiographed, or alternatively was subjected to Western blot analysis. The radiolabeled band in lane 1 corresponds in size to the band detected by a JIL-1-specific antibody on a Western blot (lane 2).
(C) The top panel shows autoradiographs of histone H3 protein. The presence of JIL-1 protein immunoprecipitated from S2 cells with Odin antiserum (Odin ip) leads to radiolabeling of histone H3 in the in vitro kinase assay. In contrast, histone H3 shows no labeling after incubation with Odin preimmune serum–immunoprecipitated protein (preimmune ip). The bottom panel shows Coomassie blue staining of the gels demonstrating that equivalent amounts of histone H3 were present in both reactions. The migration of molecular weight markers in kDa is shown for (A) and (B).
Molecular Cell 1999 4, DOI: ( /S (00) )

4. Figure 3 Imaging of Live Nuclei from JIL-1-GFP Transgenic Flies
(A) Expression of JIL-1-GFP NH2-terminal fusion protein is under the control of the hs83 promoter.
(B) JIL-1-GFP is detectable on immunoblots with Hope antiserum in transgenic flies as an additional band (arrow) compared to wild-type flies.
(C) Average projection image of ten confocal sections of two live ovarian egg chambers showing JIL-1-GFP fluorescence localized to the nuclei.
(D) Four frames from a time lapse movie of dividing nuclei in live JIL-1-GFP transgenic syncytial embryos: (D1) interphase, (D2) prophase, (D3) metaphase, and (D4) second interphase.
Molecular Cell 1999 4, DOI: ( /S (00) )

5. Figure 4 JIL-1 Expression in Salivary Gland Polytene Chromosomes
(A–C) Double labeling of female polytene chromosomes with Hope antiserum (A) and Hoechst (B) shows that JIL-1 is expressed in a banded pattern complementary to that of Hoechst. In the composite image (C), there is little overlap between JIL-1 (green) and Hoechst (red) labeling (indicated by minimal regions of yellow). (D–E) JIL-1 antibody labeling of male X chromosomes (indicated by X) is greatly enhanced as compared to autosomes. In contrast, double labeling of the preparation in (E) with Hoechst (F) shows that the male X is less intensely labeled with Hoechst as compared to autosomes. (G) The composite image of the JIL-1 (green) and Hoechst (red) labeling of the preparation in (E) and (F) shows that the complementary banding pattern is maintained on the male X (white lines). (H) Stereo pair of JIL-1-GFP fluorescence in a live female transgenic polytene nucleus. The expression of JIL-1-GFP in discrete bands is clearly discernible. (I) Average projection image of three confocal sections from a live male JIL-1-GFP transgenic polytene nucleus. The X chromosome exhibits enhanced fluorescence as compared to autosomes.
Molecular Cell 1999 4, DOI: ( /S (00) )