Journal: Molecular Biology of the Cell

Article Title: Newfound features of meiotic chromosome organization that promote efficient congression and segregation in Caenorhabditis elegans oocytes

doi: 10.1091/mbc.E22-07-0297

Figure Lengend Snippet: The ring complex has internal domains that weave through Meiosis I bivalents. (A) Meiosis I bivalents (Top) and Meiosis II chromosomes (Bottom); shown are DNA (blue), SUMO (green), and AIR-2 (red). Zooms highlight individual RCs; MI bivalents have an internal substructure (arrow; 57/57) while MII chromosomes do not (0/54 have internal RC components). (B) 3D-SIM superresolution image of an end-on Meiosis I bivalent; shown are DNA (blue), SUMO (green), and AIR-2 (red). Arrows highlight subloops within the RC. (C) 3D-SIM superresolution image of a Metaphase I-arrested monopolar spindle. This condition was generated by performing klp-18;klp-7 MCAK (RNAi) in emb-27(g48)ts worms and shifting them to the restrictive temperature. AIR-2 (red) stretches toward the plus ends of the microtubules, which reveals subloops (arrows) associated with the bivalent; subloops are apparent in the single-slice zoom (schematic shows RC organization). (D) Model for RC organization in Meiosis I. Homologous chromosomes are shown in light/dark blue and cohesin linking sisters is shown in purple. After homologs pair and a single off-centered crossover occurs, the cruciform structure condenses such that the short arms are indistinguishable from the long arms. The RC (red) forms subloops around the short arms of the bivalents as well as encircling the outside of the bivalent. Scale bars = 5 µm (full images), 1 µm (zooms).

Article Snippet: Images in Figure 1, B and C (denoted as superresolution) were acquired on an OMX 3D-SIM microscope (Applied Precision) with an Olympus 100× UPlanSApo objective (NA = 1.4) in the Center for Advanced Microscopy at Northwestern University.

Techniques: Generated

Journal: Molecular Biology of the Cell

Article Title: Cell cycle localization dynamics of mitochondrial DNA polymerase IC in African trypanosomes

doi: 10.1091/mbc.E18-02-0127

Figure Lengend Snippet: Three-dimensional SIM imaging of POLIC-PTP, TAC102, and basal bodies. TbIC-PTP cells were labeled with anti–protein A (green), anti-TAC102 (red), YL1/2 (white), and DAPI (blue). More than 20 cells were analyzed by 3D-SIM at each stage. Scale bar: 5 μm. (A) Representative images of POLIC-PTP, TAC102, and bb localization at stage IIa and stage IIb of kDNA replication. (B) Schematic model of localization of POLIC-PTP (green), TAC102 (red), and YL1/2 (gray) during stages IIa and IIb of kDNA replication. Videos of each representative image can be found in the Supplemental Material.

Article Snippet: Three-dimensional SIM (3D-SIM) was performed using a Nikon N-SIM E superresolution microscope equipped with an ORCA-Flash 4.0 sCMOS camera (Hamamatsu Photonics K.K.) and a CFI SR Apochromat TIRF 100× oil-objective (NA 1.49) lens.

Techniques: Imaging, Labeling

Journal: Molecular and Cellular Biology

Article Title: The Tumor Suppressor PML Specifically Accumulates at RPA/Rad51-Containing DNA Damage Repair Foci but Is Nonessential for DNA Damage-Induced Fibroblast Senescence

doi: 10.1128/MCB.01345-13

Figure Lengend Snippet: Superresolution imaging of PML nuclear bodies and DNA damage sites. Primary human fibroblasts (at low PD) were irradiated with 10 Gy, fixed after 90 min, and subsequently immunostained to detect the indicated proteins. 3D SIM superresolution images of single nuclei were collected using an Elyra structured-illumination microscope (Zeiss). The white boxes within the overview images are shown enlarged on the right. Each image on the right also shows the individual channels in monochrome.

Article Snippet: 3D SIM superresolution images of single nuclei were collected using an Elyra structured-illumination microscope (Zeiss).

Techniques: Imaging, Irradiation, Microscopy