Fig. 4

TAGLN mediates stiffness-regulated ovarian cancer progression and correlates with poor patient prognosis. A Knockdown of TAGLN by siRNA in SK-OV-3 cells were determined by western blot and probed for TAGLN and GAPDH. B-C Migratory (B) and invasive (C) ability of SK-OV-3 cells transfected with siControl or TAGLN siRNA, cultured on soft or stiff substrates, were evaluated by transwell assays and quantified (*P < 0.05). (D) Overexpression of TAGLN by plasmid transfection in SK-OV-3 cells were determined by western blot and probed for TAGLN and GAPDH. E-F Migratory (E) and invasive (F) ability of SK-OV-3 cells with the expression of control vector or overexpression of TAGLN protein, cultured on soft or stiff substrates, were evaluated by transwell assays and quantified (*P < 0.05, ****P < 0.0001). G-H The proliferation of SK-OV-3 cells down-regulated (G) or overexpressed TAGLN (H) was measured by EdU assay. Quantification of EdU-positive nuclei from n = 3 experiments. I Cell lysates from SK-OV-3-ip3-luc transfected with lentivirus of sh-control and sh-TAGLN were determined by western blot and probed for TAGLN and GAPDH. J-K Representative bioluminescence images (J) of mice bearing SK-OV-3-ip3-luc cells transfected with sh-control or sh-TAGLN at 4 weeks after tumor implantation. Bar graph (K) showing the quantification of normalized total photon counts in each group (n = 6, *P < 0.05). L Incidence of metastases was decreased in sh-TAGLN group (n = 6 each group). In sh-control group, two mice have peritoneal metastasis, the incidence of metastasis was 33.3 %; in sh-TAGLN group, no mouse has peritoneal metastasis, the incidence of metastasis was 0 %. M-N Kaplan–Meier analysis of PFS (M) and OS (N) of ovarian cancer patients classified by tumor TAGLN protein levels into high and low expression group. O-P PFS (O) and OS (P) in ovarian cancer patients with low and high levels of TAGLN (Affymetrix probeset 205547_s_at). PFS: progression-free survival, OS: overall survival