Journal: Physiological reports

Article Title: Effects of Lactobacillus johnsonii and Lactobacillus reuteri on gut barrier function and heat shock proteins in intestinal porcine epithelial cells.

doi: 10.14814/phy2.12355

Figure Lengend Snippet: Figure 1. Characterization of IPEC-J2 cell monolayer. (A) Light microscope image of confluent cell monolayer grown on a tissue culture flask, scale bar equals 20 lm. (B) Confocal micrograph showing a top view of IPEC-J2 monolayer grown on transwell filter for 9 days, the cell borders can be distinguished by immunofluorescent staining of tight junction protein ZO-1, scale bar equals 20 lm. (C) Top view of well- differentiated IPEC-J2 cell with microvilli observed by scanning electron microscopy, figure contains one cell from cell monolayer grown on transwell filter for 9 days, scale bar equals 5 lm. (D) Progression in transepithelial electrical resistance (TEER) values of cells grown on transwell filter for 9 days. Data are given as means (SEM) of 20 separate experiments. They were assigned to experimental treatment on day 9, respectively.

Article Snippet: Images were acquired using laser scanning confocal microscopy (Nikon C-1 with Plan ApoVC 609/1.40 oil objective; Nikon EZ-C1 software; Nikon, Amsterdam, the Netherlands).

Techniques: Light Microscopy, Staining, Electron Microscopy

Journal: Physiological reports

Article Title: Effects of Lactobacillus johnsonii and Lactobacillus reuteri on gut barrier function and heat shock proteins in intestinal porcine epithelial cells.

doi: 10.14814/phy2.12355

Figure Lengend Snippet: Figure 7. Cellular distribution of the tight junction protein ZO-1 in IPEC-J2 cells. Cell monolayers grown on transwell filters were left untreated (control) or treated with bacterial cells of ETEC (multiplicity of infection, MOI 10) alone for 3 h or pretreated with Lactobacillus spp. (MOI 100) for 6 h and then challenged by ETEC for 3 h. Monolayers stained for the tight junction protein ZO-1 (green) and nuclei stained with DAPI (blue) were detected by laser scanning confocal microscopy. The images were representatives from three separate experiments. Yellow arrows show the broken lining of ZO-1 expressions. Red circles highlight areas of cell disassociation. Scale bar equals 50 lm for all images.

Article Snippet: Images were acquired using laser scanning confocal microscopy (Nikon C-1 with Plan ApoVC 609/1.40 oil objective; Nikon EZ-C1 software; Nikon, Amsterdam, the Netherlands).

Techniques: Control, Infection, Staining, Confocal Microscopy

Journal: Journal of Biological Research

Article Title: Cortical microtubule orientation in Arabidopsis thaliana root meristematic zone depends on cell division and requires severing by katanin

doi: 10.1186/s40709-018-0082-6

Figure Lengend Snippet: Cortical microtubule orientation in the meristematic zone of untreated ( a ) and aphidicolin-treated ( b – d ) roots. Single CLSM sections through the boundary between protoderm and cortex are presented, so that in the center of each figure cortex cells can be observed. In all the images of this work the root tip is oriented towards the bottom of the page. In control root ( a ) prominent transverse orientation of cortical microtubules can be observed in interphase cells (arrowheads). At 12 h of treatment ( b ) transverse microtubule orientation persists (arrowheads), while dividing cells still exist (arrow points to a mitotic cell). After 24 h of treatment ( c ), cell divisions have ceased and cortical microtubules exhibit random orientation (cells within the brackets are cortex cells viewed at external cortical plane), which is also observed after 48 h of treatment ( d ; cells within brackets). In the latter image, vacuolation of meristematic cells in also prominent (asterisks show vacuoles). Representative decipher-graphs of microtubule alignment in meristematic cells of control ( e ), 12 h ( f ), 24 h ( g ) and 48 h ( h ) aphidicolin treatments. Decipher-graphs show that microtubules are transversely oriented (doubled-headed arrow) in the control and after 12 h of aphidicolin treatment ( e , f ), while the frequency of longitudinal and randomly oriented microtubules increased upon prolonged treatments ( g , h , respectively). Scale bar 10 μm

Article Snippet: The preparations were examined with a Nikon D-Eclipse C1 or a Zeiss LSM780 confocal laser scanning microscope (CLSM), with the appropriate filters for FITC, and micrographs were acquired with each manufacturer’s software.

Techniques: Control

Journal: Journal of Biological Research

Article Title: Cortical microtubule orientation in Arabidopsis thaliana root meristematic zone depends on cell division and requires severing by katanin

doi: 10.1186/s40709-018-0082-6

Figure Lengend Snippet: Cortical microtubule orientation at the external face of epidermal cells in the transition (i.e. under the shootward part of the lateral root cap; a , c ) and fast elongation ( b , d ) zones of aphidicolin-treated roots. All images are maximum projections of serial CLSM sections. After 24 h of treatment, microtubules are transverse (arrowheads) in the transition ( a ) and fast elongation ( b ) zone, while after 48 h of treatment they appear randomly oriented in both zones ( c , d ). Representative decipher-graphs of microtubule alignment in transition and elongation zone cells of control roots ( e ), and after 24 h ( f ) and 48 h ( g ) of aphidicolin treatment. Decipher-graphs show that microtubules are transversely oriented (doubled-headed arrow) in the control and after 24 h of aphidicolin treatment ( e and f , respectively), while the frequency of longitudinal and/or randomly oriented microtubules increased after 48 h of treatment ( g ). Scale bar 10 μm

Article Snippet: The preparations were examined with a Nikon D-Eclipse C1 or a Zeiss LSM780 confocal laser scanning microscope (CLSM), with the appropriate filters for FITC, and micrographs were acquired with each manufacturer’s software.

Techniques: Control

Journal: Journal of Biological Research

Article Title: Cortical microtubule orientation in Arabidopsis thaliana root meristematic zone depends on cell division and requires severing by katanin

doi: 10.1186/s40709-018-0082-6

Figure Lengend Snippet: Cortical microtubule orientation in cortex cells of wild-type ( a , e ) and p60-katanin mutants ( b – d , f – h ). All high magnification images ( e – h ) are maximum projections of serial CLSM sections through the cortical cytoplasm. Cortex cell files, derived from the areas defined by rectangular frames on low magnification images ( a – d ), were released after application of gentle pressure on the whole mount root specimens. Fairly transverse cortical microtubules can be observed in Col-0 cells ( e ). In ktn1 - 2 ( f ), lue1 ( g ) and fra2 ( h ), though a general transverse pattern exists, several microtubules exhibit random orientation, while foci of microtubule convergence can be discerned as well. Representative decipher-graphs of microtubule alignment in meristematic cortex cells of Col-0 ( i ), ktn1 - 2 ( j ), lue1 ( k ) and fra2 ( l ). Generally, transverse microtubule orientation prevailed in all the cases (doubled-headed arrow). However, in the katanin mutants an increase in the frequency of longitudinal and/or randomly oriented microtubules was noticed. Scale bars 50 μm ( a – d ), 10 μm ( e – h )

Article Snippet: The preparations were examined with a Nikon D-Eclipse C1 or a Zeiss LSM780 confocal laser scanning microscope (CLSM), with the appropriate filters for FITC, and micrographs were acquired with each manufacturer’s software.

Techniques: Derivative Assay, Gentle

Journal: Biochemical Pharmacology

Article Title: HDAC 3-selective inhibitor RGFP966 demonstrates anti-inflammatory properties in RAW 264.7 macrophages and mouse precision-cut lung slices by attenuating NF-κB p65 transcriptional activity

doi: 10.1016/j.bcp.2016.03.010

Figure Lengend Snippet: RGFP966 does not affect the localization of HDAC 3 and HDAC 3 – NF-κB p65 interaction in LPS/IFNγ-stimulated RAW 264.7 macrophages. After 20 h incubation with RGFP966 followed by 1 h LPS/IFNγ stimulation, RAW 264.7 macrophages were prepared for immunofluorescence microscopy (A). The green signal represents NF-kB p65 protein, while the blue signal visualizes the Hoechst-stained nuclei. RGFP966 did not affect the nuclear localization of HDACs 1–3 compared to (vehicle-treated) control group. The presented data set shows representative images of 4 independent experiments. Images of the cells were taken using confocal laser scanning microscopy and all images were taken with identical instrumental conditions, original magnification 630×. In addition, the effect of RGFP966 on HDAC 1–3 localization in LPS/IFNγ-stimulated RAW 264.7 macrophages was analyzed by immunoblotting cell fractions (data not shown) and quantified by densitometric analysis (B). PARP-1 and β-actin were used as internal controls. Control (vehicle-treated) cells were set at 100%. Data are presented as mean values ± SD of 3–4 independent experiments. The HDAC 3 – NF-κB p65 interaction in LPS/IFNγ-stimulated RGFP966-treated RAW 264.7 macrophages was investigated by immunoprecipitation of NF-κB p65 followed by immunoblotting for HDAC 3, which was quantified by densitometric analysis (C). Protein levels were normalized against NF-κB p65. Data are presented as mean values ± SD of 3 independent experiments and a representative blot is shown. * p < 0.05 compared to vehicle. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Subsequently, lysates were incubated with 50 μl of Protein A/G PLUS-Agarose Immunoprecipitation Reagent (sc-2003; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at constant rotation for 6 h at 4 °C.

Techniques: Incubation, Immunofluorescence, Microscopy, Staining, Control, Confocal Laser Scanning Microscopy, Western Blot, Immunoprecipitation