FFSS increases 14-3-3θ expression and its interaction with Cx43 and integrin α5. (A) PC12 cell lysates (lane 1) or lysates of MLO-Y4 cells subjected to fluid flow for 0 (lane 2), 0.5 (lane 3), 2 (lane 4), and 24 (lane 5) hours were immunoblotted with anti-14-3-3θ or GAPDH antibody. Band intensity was quantified and the normalized ratio of 14-3-3θ to GAPDH was calculated (right panel). **P<0.01 for 2 hours versus 0 hours; n = 3. (B,C) Lysates of MLO-Y4 cells subjected to FFSS for 2 hours (lane 3) or non-fluid flow (lanes 1 and 2) were immunoprecipitated with anti-14-3-3θ antibody. Preloaded lysates (input) or eluates from beads were immunoblotted with anti-Cx43 (B) or anti-integrin α5 (C) antibody. Band intensity of Cx43 was quantified and the normalized ratio of eluates from beads (IP) to preloaded (input) was calculated (lower panel). In B,C, *P<0.05 for FF versus non-FF (C); n = 3.