Figure 6: The direct repression of RRAS by miR-449 affects multiciliogenesis and apical actin cytoskeleton reorganization in HAECs. | Nature Communications

Figure 6: The direct repression of RRAS by miR-449 affects multiciliogenesis and apical actin cytoskeleton reorganization in HAECs.

From: miR-34/449 control apical actin network formation during multiciliogenesis through small GTPase pathways

Figure 6

(a) Differentiating HAECs were chronically treated with negative oligonucleotide (CTR-Neg, a1–3) or with an oligonucleotide protecting the miR-449-binding site on RRAS (PO-RRAS, a4–6) and stained at LC stage for F-actin (a1,4), ezrin (a2,5) and acetylated tubulin (Ac. Tub., a3,6). (b) Alternatively, differentiating HAECs were transfected at seeding time with si-RRAS and stained at LC stage for F-actin (in red, c1) and motile cilia (Ac. Tub. in magenta, c2). (c) The histogram indicates the number of apical ezrin-positive cells (in green) and MCCs (in magenta) normalized in percentage from control. Protecting the RRAS mRNA from interaction with miR-449 leads to defects in apical actin reorganization together with a decrease in MCC differentiation similar to that observed after inhibition of miR-449 activity. All data are means±s.d. from at least three independent experiments (***P<0.001; Student’s t-test).

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