1. Global Transcriptional Repression in C
Global Transcriptional Repression in C. elegans Germline Precursors by Regulated Sequestration of TAF-4 
Tugba Guven-Ozkan, Yuichi Nishi, Scott M. Robertson, Rueyling Lin 
Cell 
Volume 135, Issue 1, Pages (October 2008)
DOI: /j.cell
Copyright © 2008 Elsevier Inc. Terms and Conditions

2. Figure 1
Schematic of C. elegans Hermaphrodite Gonad and Early Embryonic Divisions
(A) Oocyte development and maturation schematized in one of the two gonad arms. Germ nuclei (white circles) are in a syncitial cytoplasm of the gonad arm before cellularization to form oocytes. The oocyte adjacent to the spermatheca (−1) undergoes maturation and is then immediately ovulated and fertilized.
(B) Early embryonic divisions, highlighting the germline precursors P0 to P4 (red) and their somatic sisters (green). Left, schematic; right, differential interference contrast (DIC) images. Embryo images in this and all subsequent figures are oriented with the anterior pole to the left. The scale bar represents 10 μm.
Cell  , DOI: ( /j.cell )
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3. Figure 2
OMA-1 and OMA-2 Are Required for Nuclear Enrichment of GFP::TAF-4 in One-Cell Embryos
(A) DIC and GFP fluorescence micrographs of GFP::TAF-4-expressing embryos (first two columns), anti-TAF-4 staining of wild-type embryos (third column), and anti-GFP and anti-OMA-1 double staining of GFP::TAF-4-expressing embryos (last two columns). DC, pronuclei decondensation; PM, pronuclei meeting. Arrowhead points to pronuclei in one-cell embryos.
(B) DIC and GFP fluorescence micrographs of animals expressing various GFP-fused transgenic proteins, as indicated below the DIC image. Embryos in the uterus arrange in a developmental progression from youngest (left) to oldest (right). −1, maturing oocyte.
(C) GFP fluorescence in fixed embryos from transgenic animals expressing GFP::TAF-4 either without (left) or after (right) oma-1(RNAi);oma-2(RNAi). White arrowheads, two-cell embryos. The two-cell and two one-cell embryos not circled in non-RNAi samples are dividing.
One- and 12-cell embryos are outlined in dashed and solid white lines in (B) and (C). Scale bars represent 25 μm.
Cell  , DOI: ( /j.cell )
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4. Figure 3
OMA-1 and OMA-2 Are Required for Transcriptional Repression in Germline Blastomeres
(A) Immunofluorescence micrographs of anti-Ser2P staining in wild-type (non-RNAi), pie-1(RNAi), oma-1(RNAi);oma-2(RNAi), and oma-1(te33);oma-2(RNAi);pie-1(RNAi) embryos. The background cytoplasmic signals in one-cell (1C) and two-cell (2C) non-RNAi and pie-1(RNAi) embryos were enhanced to reveal the outline of embryos. 4C, four-cell embryos.
(B) In situ hybridization of the vet-5 gene. For each embryo, DIC and DAPI images are shown. Open arrowheads point to pronuclei in 1C, nuclei in 2C, and the germline nucleus in 4C embryos, respectively.
(C) anti-Ser2P in the gonad of wild-type or oma-1(te33);oma-2(te51) animals. Oocyte nuclei are indicated by arrows. sp, spermatheca.
(D) Immunofluorescence micrographs of anti-GFP and anti-Ser2P double staining in embryos expressing GFP::PIE-1. Left two columns, non-RNAi; right two columns, oma-1(RNAi);oma-2(RNAi).
Scale bars represent 10 μm.
Cell  , DOI: ( /j.cell )
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5. Figure 4 OMA-1 Interacts with TAF-4 Histone Fold Domain
(A and B) Deletion analysis to map the TAF-4 (A) and OMA-1 (B) interaction domains. HFD, histone fold domain; TAFH, TAF homology domain; ZF, TIS11 zinc finger. Numbers to the left of each construct indicate amino acid positions. Red dashed lines demarcate those regions that are necessary and sufficient for the interaction.
(C) Top: Sequence alignment of the HFD from various TAF-12 proteins compared to OMA-1 and OMA-2 sequences including the TAF-4 interacting domain. Bottom: Sequence alignment of various TAF-4 proteins. Underlined in red are the regions necessary and sufficient for interaction. Hs, human; Dm, D. melanogaster; Sc, S. cerevisiae; Ce, worm. Yellow highlighting indicates residues located in the hydrophobic core of the TAF-4/TAF-12 heterodimer (Werten et al., 2002). α, alpha helix; L, loop. Amino acids conserved among TAF proteins are highlighted. Blue, hydrophobic; red, charged; green, small.
(D) Immunofluorescence micrographs of anti-GFP and anti-Ser2P double staining or anti-TAF-4 staining in oma-1(te33) embryos expressing OMA-1 Δ46-80::GFP that are treated with either no RNAi or oma-2(RNAi). The scale bar represents 10 μm.
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6. Figure 5 OMA-1 Competes with TAF-12 for TAF-4 Binding
(A) GFP fluorescence in the gonad (dashed line) and embryos of transgenic worms expressing GFP::TAF-4, GFP::TAF-4ΔHFD, or GFP::TAF-4 with taf-12(RNAi).
(B) HEK293 cells expressing TAF-4 (green) alone, TAF-4+TAF-12 (magenta), TAF-4+MYR::OMA-1-N (red), or TAF-4+TAF-12+MYR::OMA-1-N.
(C) TAF-4ΔHFD expressed alone, with TAF-12, or with Myr::OMA-1-N.
The scale bar represents 25 μm.
Cell  , DOI: ( /j.cell )
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7. Figure 6 Interaction between OMA-1 and TAF-4 Is Regulated by MBK-2
(A) Top three rows: GFP fluorescence in utero of embryos expressing GFP::TAF-4 in genetic backgrounds listed to the left. Bottom row: persistence of GFP::OMA-1 fluorescence after mbk-2(RNAi). Arrows mark one-cell embryos.
(B) Immunofluorescence micrographs of anti-Ser2P and anti-GFP double staining in one-cell embryos expressing GFP::TAF-4 either with or without mbk-2(RNAi).
(C) Confocal immunofluorescence micrographs of Myr::OMA-1-FL, GFP::TAF-4, DAPI, and all three (merged) in HEK239 cells. Proteins expressed from transfected constructs are indicated on the left. KD, kinase dead. The localization of Myr-OMA-1-FL (cytoplasmic membrane and cytoplasmic aggregates that were observed when high levels of Myr-tagged proteins were expressed) is not affected by MBK-2 activity.
Scale bars represent 25 μm (A) and 10 μm (B).
Cell  , DOI: ( /j.cell )
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8. Figure 7
Ectopic OMA-1 in cul-1-Depleted Embryos Can Repress Transcription
(A) GFP fluorescence and corresponding DIC images of four-cell embryos expressing GFP::TAF-4 and embryos in utero expressing GFP::OMA-1 with or without cul-1(RNAi). Arrow, one-cell embryo; arrowhead, four-cell embryo.
(B) Immunofluorescence micrographs of anti-Ser2P and anti-OMA-1 double staining in four-cell embryos of the following backgrounds: no RNAi, cul-1(RNAi), cul-1(RNAi);oma-1(te33);oma-2(RNAi), cul-1(RNAi);oma-1(te33);oma-1(RNAi), pie-1(RNAi);cul-1(RNAi), or pie-1(RNAi);oma-2(RNAi).
Scale bars represent 20 μm (A) and 10 μm (B).
Cell  , DOI: ( /j.cell )
Copyright © 2008 Elsevier Inc. Terms and Conditions