Journal: Integrative Organismal Biology

Article Title: Regeneration and Musculature in Halved Cassiopea xamachana Ephyrae

doi: 10.1093/iob/obaf030

Figure Lengend Snippet: Stereo microscope pictures of C. xamachana ephyrae showing growth of intact and bisected ephyrae. (A) Trajectory of growth in the original ephyra with photographs at days 0, 7, 14, 21, and 28. (B) Visual progress of two fragments with circle back comparison to original medusa, all are on the same scale (see scale bar). Locations of rhopalia are pinpointed with black dots, edges of the bell and oral arms are outlined in black. See description of one ephyra regeneration and development in .

Article Snippet: Ephyrae were then washed in PBS, mounted on a slide and imaged using a confocal microscope (ImageXpress Pico, Molecular devices).

Techniques: Microscopy, Comparison

Journal: Integrative Organismal Biology

Article Title: Regeneration and Musculature in Halved Cassiopea xamachana Ephyrae

doi: 10.1093/iob/obaf030

Figure Lengend Snippet: Confocal microscope images of actin-stained regenerating ephyrae at timepoints 0 and 24 h after bisection. (A) T (time) = 0 h post amputation (hpa); (B) T = 24 hpa after cut; (C) Close up of the wound site at T = 24 hpa. Scale bar A, B = 1 mm. Scale bar C = 50 µm. See for T = 48 hpa and T = 72 hpa.

Article Snippet: Ephyrae were then washed in PBS, mounted on a slide and imaged using a confocal microscope (ImageXpress Pico, Molecular devices).

Techniques: Microscopy, Staining

Journal: Cells

Article Title: ‘Shared-Hook’ and ‘Changed-Hook’ Binding Activities of Herpesviral Core Nuclear Egress Complexes Identified by Random Mutagenesis

doi: 10.3390/cells11244030

Figure Lengend Snippet: Qualitative and quantitative analyses of HCMV AD169-GFP ΔUL53 virus reconstitution on the different recombinant HFF cell populations. HFFs that express pUL53, pUL53-Flag or pUL53::sHook1-Flag were transfected with the infectious bacterial artificial chromosome of HCMV ΔUL53. ( A ) Images of the GFP signals and the respective brightfield illuminations were taken at indicated time points; scale bar in panel A, picture 1 marks 500 μm. ( B ) Quantitative analysis of fluorescence-positive cells was achieved with automated counting by the CellReporterXpress ® software (Molecular Devices LLC, San Jose, CA, USA). Counting parameters for positive nuclei were set to an intensity of at least 100, a minimal width of 7 and a maximal width of 30. Measurements were performed in biological sextuplicates per cell population of 5.05% area of the well and mean values ± SD are given. ( C ) qPCR-based assay for the determination of viral genome equivalents referring to the respective recombinant HFF populations. Viral supernatants were harvested at day 24 p.t. and subjected to IE1-specific qPCR. Calculations were performed in biological sextuplicates per cell population; mean values ± SD are shown. ( B , C ) Statistical analysis was performed using an ordinary two-way ANOVA and post-hoc Tukey correction; ****, p < 0.0001; **, p < 0.01; *, p < 0.1; ns, not significant; no significant differences were noted for values between 1–20 d p.t. ( B ). ( D ) Wb-based expression analysis of the conditionally expressing HFF populations. Recombinant protein expression in the three HFF populations of HFF-UL53, HFF-UL53-Flag and HFF-UL53::sHook1-Flag cells was either uninduced (-) or induced (500 ng/mL Dox). Cells were either infected with HCMV ΔUL53 or remained mock-infected. At 24 h p.t., cells were harvested and lysed. Total lysate samples were subjected to standard Wb analysis using tag-specific or protein-specific monoclonal antibodies as indicated.

Article Snippet: For measurement of HCMV AD169-GFP ΔUL53-positive cells, signals were counted with the ImageXpress ® Pico device (Molecular Devices LLC, San Jose, CA, USA) using CellReporterXpress ® software (version 2.9.3.1183, Molecular Devices LLC) by the system-integrated cell count assay.

Techniques: Virus, Recombinant, Transfection, Fluorescence, Software, Expressing, Infection, Bioprocessing

Journal: Cells

Article Title: ‘Shared-Hook’ and ‘Changed-Hook’ Binding Activities of Herpesviral Core Nuclear Egress Complexes Identified by Random Mutagenesis

doi: 10.3390/cells11244030

Figure Lengend Snippet: Viral replication kinetics of HCMV ΔUL53 and its revertant (HCMV Rev) determined by quantitation of GFP-positive cells and HCMV-specific qPCR on the different recombinant HFF populations. 80,000 inducibly expressing HFFs in 24-well plates were infected with HCMV ΔUL53 or HCMV Rev at a viral dose of 5 × 10 6 genome copies. pUL53, pUL53-Flag or pUL53::sHook1-Flag protein expression was either Dox-induced (+Dox) or remained non-induced (−Dox). ( A ) The number of HCMV-infected cells was measured by detection of GFP signal-positive cells at indicated time points with the CellReporterXpress ® software using the ImageXpress ® Pico device. Values represent 25.04 % of the area of a well and are given as a mean value ± SD of two independently infected wells. ( B ) Viral supernatants were harvested at indicated time points and viral genome equivalents were determined by qPCR. Each value represents the mean ± SD of two independent biological replicates, each measured twice. ( A , B ) Statistical analysis was performed using an ordinary two-way ANOVA and post-hoc Sidak correction; ****, p < 0.0001; ***, p < 0.001; *, p < 0.1; ns, not significant; no significant differences were noted for values between 1–13 d p.i. ( A ) or 1–11 d p.i. ( B ), respectively.

Article Snippet: For measurement of HCMV AD169-GFP ΔUL53-positive cells, signals were counted with the ImageXpress ® Pico device (Molecular Devices LLC, San Jose, CA, USA) using CellReporterXpress ® software (version 2.9.3.1183, Molecular Devices LLC) by the system-integrated cell count assay.

Techniques: Quantitation Assay, Recombinant, Expressing, Infection, Software