Journal: Micromachines

Article Title: A Microfluidic-Based Cell-Stretching Culture Device That Allows for Easy Preparation of Slides for Observation with High-Magnification Objective Lenses.

doi: 10.3390/mi16010093

Figure Lengend Snippet: Figure 5. Effects of cyclic stretching on integrinβ1 and vinculin localization in OP9 cells. Fluorescence images of integrinβ1 (green), f-actin (red), and vinculin (gray) in OP9 cells cultured on the device after 72 h under static conditions followed by 24 h under (A) static or (B) cyclic stretching conditions (0.5 Hz) photographed with a 100× oil immersion objective lens. Confocal images were captured every 0.48 µm along the z-axis to create z-stacks (14 slices) for maximum-intensity projection (scanning zoom 2×). The right figure shows the fluorescence intensity–distance profiles of integrinβ1 (green), f-actin (red), vinculin (gray), and nuclei (blue) on lines (a–d) drawn on the microscope image (MIP).

Article Snippet: For integrinβ1, vinculin, f-actin, and nuclear staining, 0.5% BSA was added with anti-integrinβ1 antibody (0.4 μg/mL; SC-19656, Santa Cruz Biotechnology, Dallas, TX, USA), anti-vinculin antibody (4 μg/mL; 26520-1-AP, Proteintech, Rosemont, IL, USA), and rhodamine phalloidin (1.65 μM; R4159, Thermo Fisher Scientific) and incubated overnight at 4 ◦C.

Techniques: Fluorescence, Cell Culture, Microscopy

Journal:

Article Title: Coxsackie and adenovirus receptor (CAR) is a product of Sertoli and germ cells in rat testes which is localized at the Sertoli-Sertoli and Sertoli-germ cell interface *

doi: 10.1016/j.yexcr.2007.01.017

Figure Lengend Snippet: Sources of antibodies and their working dilutions used for different experiments in this report

Article Snippet: Lot No. Use b Working dilution c Santa Cruz Biotechnologies (Santa Cruz,CA) CAR Rabbit sc-15405 J1304 IP IB 1:200 IF 1:50 IHC 1:100 CAR Goat sc-10313 F0304 IHC 1:100 CAR Mouse, monoclonal sc-32795 E2605 IB 1:200 Occludin Rabbit sc-5562 G2803 IB 1:200 Jam-C Goat sc-23005 L1704 IF 1:50 Actin Goat sc-1616 J0104 IB 1:200 CAR Goat sc-10313 F0304 IP α-catenin Rabbit sc-7894 A2705 IP β - catenin Rabbit sc-7199 F0204 IP γ - catenin Rabbit sc-7900 J139 IP Vinculin Rabbit sc-5573 I 2204 IP α-actinin Goat sc-7453 D292 IP FAK Rabbit sc-558 D1806 IP c-Src Mouse, monoclonal sc-8056 F3006 IF 1:100 Upstate Biotechnology (Lake Placid,NY) PI-3kinase Rabbit 06497 25006 IP p130 cas Rabbit 06500 19950 IP Src,CT,clone NL19 Rabbit, Monoclonal 05–772 26812 IP IB 1:1000 BD Biosciences (San Jose, CA) Integrinβ1 Mouse, monoclonal 610468 53855 IP Espin Mouse, monoclonal 611656 32291 IF 1:100 Zymed Laboratories Inc. (South San Francisco, CA) N-Cadherin Mouse, monoclonal 33–3900 50393487 IF 1:50 ZO-1 FITC conjugate Mouse, monoclonal 33–91111 60102882 IF 1:50 Jam-A Rabbit 36–1700 40890360 IB 1:200 Goat anti-mouse FITC conjugate Goat 81–6511 40397219 IF 1:50 Goat anti-rabbit Cy3 conjugate Goat 81–6115 51001169 IF 1:50 Chemicon (Temecula, CA) Donkey anti-mouse FITC conjugate Donkey AP192F 508006630 IF 1:100 Donkey anti-rabbit Cy3 conjugate Donkey AP182C 24010618 IF 1:100 Donkey anti-goat FITC conjugate Donkey AP180F 24011222 IF 1:100 Open in a separate window a All primary antibodies used in this report were polyclonal antibodiesunless otherwise specified.

Techniques:

Journal:

Article Title: Coxsackie and adenovirus receptor (CAR) is a product of Sertoli and germ cells in rat testes which is localized at the Sertoli-Sertoli and Sertoli-germ cell interface *

doi: 10.1016/j.yexcr.2007.01.017

Figure Lengend Snippet: A: Immunofluorescent staining of rat testes sections shows co-localization of CAR and JAM-C in vivo. Both proteins were found to be distributed on round spermatids and were confined to the heads of elongating/elongate spermatids. Scale bar = 5 μm B: Rat germ cells were isolated by mechanical procedure (without glass wool), plated on poly-L-lysine coated coverslips and permeabilized before staining. (a) Localization of CAR on germ cells: Immunofluorescent staining was performed on the slides with rabbit anti-CAR (H-300) polyclonal antibody, followed by incubation with Cy3 conjugated donkey anti-rabbit IgG. Scale bar = 25 μm, which also applies to b & c. (b) Germ cells are visualized under light microscope. (c) Merged image of CAR and DAPI staining of nuclei. (i–iv) Magnified views of individual germ cells at different development stages. (i) spermatocyte (ii) round spermatid (iii) elongate spermatid (step17–18) (iv) elongating spermatid (step 9–10). Scale bar = 5 μm in (i), which applies to (i–iv). C: Immunofluorescent micrographs of a stage VIII seminiferous tubule. CAR (red) was stained with a rabbit anti-CAR (H-300) polyclonal antibody and espin (green) was stained with a mouse-anti espin monoclonal antibody. Merged image (c) shows that CAR and espin were colocalized to the apical ES site in the seminiferous epithelium. Scale bar= 50 μm.

Article Snippet: Lot No. Use b Working dilution c Santa Cruz Biotechnologies (Santa Cruz,CA) CAR Rabbit sc-15405 J1304 IP IB 1:200 IF 1:50 IHC 1:100 CAR Goat sc-10313 F0304 IHC 1:100 CAR Mouse, monoclonal sc-32795 E2605 IB 1:200 Occludin Rabbit sc-5562 G2803 IB 1:200 Jam-C Goat sc-23005 L1704 IF 1:50 Actin Goat sc-1616 J0104 IB 1:200 CAR Goat sc-10313 F0304 IP α-catenin Rabbit sc-7894 A2705 IP β - catenin Rabbit sc-7199 F0204 IP γ - catenin Rabbit sc-7900 J139 IP Vinculin Rabbit sc-5573 I 2204 IP α-actinin Goat sc-7453 D292 IP FAK Rabbit sc-558 D1806 IP c-Src Mouse, monoclonal sc-8056 F3006 IF 1:100 Upstate Biotechnology (Lake Placid,NY) PI-3kinase Rabbit 06497 25006 IP p130 cas Rabbit 06500 19950 IP Src,CT,clone NL19 Rabbit, Monoclonal 05–772 26812 IP IB 1:1000 BD Biosciences (San Jose, CA) Integrinβ1 Mouse, monoclonal 610468 53855 IP Espin Mouse, monoclonal 611656 32291 IF 1:100 Zymed Laboratories Inc. (South San Francisco, CA) N-Cadherin Mouse, monoclonal 33–3900 50393487 IF 1:50 ZO-1 FITC conjugate Mouse, monoclonal 33–91111 60102882 IF 1:50 Jam-A Rabbit 36–1700 40890360 IB 1:200 Goat anti-mouse FITC conjugate Goat 81–6511 40397219 IF 1:50 Goat anti-rabbit Cy3 conjugate Goat 81–6115 51001169 IF 1:50 Chemicon (Temecula, CA) Donkey anti-mouse FITC conjugate Donkey AP192F 508006630 IF 1:100 Donkey anti-rabbit Cy3 conjugate Donkey AP182C 24010618 IF 1:100 Donkey anti-goat FITC conjugate Donkey AP180F 24011222 IF 1:100 Open in a separate window a All primary antibodies used in this report were polyclonal antibodiesunless otherwise specified.

Techniques: Staining, In Vivo, Isolation, Incubation, Light Microscopy