Figure 2
Figure 2. Effects of Gna13 on cellular adhesion in vivo and in vitro. (A) Representative histogram of F-actin expression in GC B cells (upper) or FO B cells (lower) from AID-Cre+ Gna13 mice. (B) MFI for F-actin expression in AID-Cre+ Gna13 mice as measured by flow cytometric analysis in spleen. Dots represent individual animals. Data are representative of 3 experiments. (C) Western blot showing levels of active (guanosine triphosphate–bound) RhoA, total RhoA, and β-actin in B-cell lysates from spleen (left) and Peyer patches (right) of Mb1-Cre+ Gna13 mice. Data are representative of 3 experiments. (D) Confocal images of Raji cell line stably transfected with lentiviral constructs: vector (N1), wild-type GNA13, dominant-negative (DN) GNA13-DN (G225A), and constitutively active (CA) GNA13-CA (Q226L). Top row: differential interference contrast; bottom row: phosphotyrosine (pTyr) staining in red. Images are representative examples of 3 experiments. Cells were imaged with a Zeiss LSM510 confocal microscope with a 63×/1.4 NA (numerical aperture) oil immersion lens. (E) Representative histogram and quantitation of pTyr expression by immunofluorescence in Raji cells transfected with constructs described in panel D. Values are quantitated as a percentage of N1 control. Results are representative of 3 experiments. (F) Representative histograms of F-actin expression in Raji cell line expressing the constructs described in panel D. Left: nontreated (N/T); right: in the presence of interleukin-6 (IL-6; 20 ng/mL). MFI, mean fluorescence intensity.

Effects of Gna13 on cellular adhesion in vivo and in vitro. (A) Representative histogram of F-actin expression in GC B cells (upper) or FO B cells (lower) from AID-Cre+Gna13 mice. (B) MFI for F-actin expression in AID-Cre+Gna13 mice as measured by flow cytometric analysis in spleen. Dots represent individual animals. Data are representative of 3 experiments. (C) Western blot showing levels of active (guanosine triphosphate–bound) RhoA, total RhoA, and β-actin in B-cell lysates from spleen (left) and Peyer patches (right) of Mb1-Cre+Gna13 mice. Data are representative of 3 experiments. (D) Confocal images of Raji cell line stably transfected with lentiviral constructs: vector (N1), wild-type GNA13, dominant-negative (DN) GNA13-DN (G225A), and constitutively active (CA) GNA13-CA (Q226L). Top row: differential interference contrast; bottom row: phosphotyrosine (pTyr) staining in red. Images are representative examples of 3 experiments. Cells were imaged with a Zeiss LSM510 confocal microscope with a 63×/1.4 NA (numerical aperture) oil immersion lens. (E) Representative histogram and quantitation of pTyr expression by immunofluorescence in Raji cells transfected with constructs described in panel D. Values are quantitated as a percentage of N1 control. Results are representative of 3 experiments. (F) Representative histograms of F-actin expression in Raji cell line expressing the constructs described in panel D. Left: nontreated (N/T); right: in the presence of interleukin-6 (IL-6; 20 ng/mL). MFI, mean fluorescence intensity.

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