Fig. 3
From: Hypoxia alters the recruitment of tropomyosins into the actin stress fibres of neuroblastoma cells
Hypoxia alters the expression and localization of HMW tropomyosins. a Normoxic (N) and hypoxic (H) SH-EP lysates were separated by SDS-PAGE. Representative Western blot stained with α9d antibody, which detects HMW tropomyosin isoforms Tm1, Tm2 and Tm3. Actin used loading control. Densitometry values were normalized to normoxic levels (dashed line) for Tm1 (b), Tm2 (c) and Tm3 (d) expression. Results are mean ± SEM; n = 3–4, * P <0.05, ** P <0.01 (t-test). e-l SH-EP cells were grown for 72 h ± hypoxia, fixed, permeabilized and incubated with Tm311 and CGβ6 antibodies to detect HMW isoforms Tm1/2/3 (e-h) and Tm2/3 (e-h), respectively. Cells then incubated with Alexa555-tagged goat anti-mouse and single z-plane images obtained by confocal microscopy. Hypoxia increased filamentous organization of Tm1/2/3 (f vs. h) and Tm2/3 (j vs. l) compared to normoxic control cells. Independent experiments (Tm311, n = 2; CGβ6, n = 5). Scale bar = 50 μm, 10 μm in inset