<i>TUG1</i> is not important for basal cell turnover, barrier or mitochondrial function, migration and monocyte adhesion.

Abstract

(A)–(G) HUVECs were transfected with two LNA GapmeRs against TUG1—LNA TUG1_1 and LNA TUG1_2 –and LNA Ctrl (10 nM) and (A) expression levels were measured after 48 hours by RT-qPCR. Expression is relative to GAPDH (n = 4; SEM; RM one-way ANOVA with Greenhouse-Geisser correction and Sidak multiple comparison test). (B) Relative cell growth determined from cell count at 0 h, 24 h, 48 h and 72 h (n = 3; SEM; RM Two-way ANOVA with Tuckey multiple comparison test). (C) Caspase-3/7 activity was measured by determination of fluorescence with ELISA plate reader (n = 3; SEM; One-way ANOVA with Holm-Sidak correction). Staurosporine was taken along as a postive control. (D) Cell-cell interactions (Rb) and cell-matrix-interactions (α) were measured by Electric Cell Impedance Sensing (ECIS; n = 3; SEM; Kruskal-Wallis-test with Dunn´s correction). (E) Determination of re-establishment of monolayer after wounding using ECIS (n = 3; SEM; One-way ANOVA with Holm-Sidak multiple comparison test). (F) Seahorse mitochondrial stress test assessing multiple mitochondrial characteristics via measurement of changes in Oxygen Consumption Rate (OCR) after serial injection of Oligomycin, Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) and Rotenone A/Antimycin (n = 3; SEM; One-way ANOVA with Holm-Sidak multiple comparison test. One representative experiment displaying the changes of OCR throughout the progress of the Seahorse mitochondrial stress test assay. (G) Assessment of monocyte adhesion with and without TNF-α stimulation. (n = 3; SEM; Two-way ANOVA with Tuckey multiple comparison test).</p