Xenbase Image ID: 150237

Figure 5. Cellular specificity of expression of ARHGAP1, ARHGDIB and R-Ras during MCC differentiation.(a,b) Fluorescence immunocytochemistry experiments on cytospins from dissociated HAECs (a1–6) and primary HAEC cultures at LC step (b) illustrate the cell-specific expression of human ARHGDIB (in green, a5–6,b) and R-Ras proteins (in green, a3,4). Basal and ciliated cells are CD151+ (in red, a1,3,5) or acetylated tubulin+ (in magenta a2,4,6 and b), respectively. Panel a2 is a magnification of an isolated acetylated tubulin-positive MCC. HAECs are stained for nuclei with 4,6-diamidino-2-phenylindole (DAPI; in blue, a1–6), F-Actin with phalloidin (in red, b). Expression levels of miR-449a (c) and of ARHGAP1, ARHGDIB and R-Ras proteins (d) during HAEC differentiation. MiR-449 levels are normalized with RNU44 (c) and HSP60 was used as a loading control (d). (e) FISH analysis of rras (e1,2) and arhgap1 (e3,4) mRNA on sections of Xenopus embryonic epidermis at stages 16 and 19 (e1,2) or at stages 17 and 20 (e3,4). MCC precursors are positive for α-tubulin (magenta; e1–4). DAPI staining is in blue and white dotted lines indicate the surface of the outer layer. (f) Real-time RT–PCR of rras transcripts in Xenopus epidermis performed at each corresponding stages indicted in the figure. (g) Transcript levels of rras decrease concomitantly with the induction of miR-449 expression. Transcript levels of rras and miR-449 were normalized and compared with their respective values obtained in stage 13. Transcript levels of rras were normalized against Odc transcript as an internal control and miR-449 expression was normalized with U2 as an internal control. Data represent the mean and s.d. of three independent experiments (***P<0.001, **P<0.01 and *P<0.05; Student's t-test). Image published in: Chevalier B et al. (2015) Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

Return to previous page