Journal: Cellular Signalling

Article Title: Deletion of inositol hexakisphosphate kinase 1 (IP6K1) reduces cell migration and invasion, conferring protection from aerodigestive tract carcinoma in mice

doi: 10.1016/j.cellsig.2016.04.011

Figure Lengend Snippet: IP6K1 modulates the invasive potential of cancer cells. (A) HCT116 cells stably expressing NT or sh IP6K1–1 were allowed to invade a matrigel matrix to move towards a high-serum gradient for 24 h. Representative images show cells that migrated through the gel to the other side of the membrane, visualized by staining with DAPI. Scale bars represent 50 μm. (B) Quantification of (A); bar graphs show the number of invaded cells normalized to the NT control. Data are mean ± SEM from five independent experiments, and were analyzed by a one sample t -test. (C) Immunoblot analysis of epithelial marker E-Cadherin in HCT116 cells expressing NT or sh IP6K1–1 . (D) Quantification of (C); levels of the epithelial marker E-cadherin are indicated as a ratio with respect to the levels of GAPDH which was the loading control. Data represents mean ± SEM from three independent experiments and was analyzed using a two tailed unpaired Student's t -test. **P ≤ 0.01; ***P ≤ 0.001.

Article Snippet: Primary antibodies used for immunoblot analysis were obtained from the following sources: Rabbit anti-IP6K1 (HPA040825, Sigma-Aldrich), goat anti-IP6K1 (sc-10419, Santa Cruz Biotechnology), rabbit anti-phosphoFAK (Tyr397) (3283, Cell Signaling Technology), rabbit anti-FAK (3285, Cell Signaling Technology), rabbit anti-phosphoPaxillin (Tyr118) (2541, Cell Signaling Technology), mouse anti-Paxillin (610051, BD Biosciences), mouse anti-actin (ab3280, Abcam), mouse anti-GAPDH (G8795, Sigma-Aldrich), mouse anti-E Cadherin (14472S, Cell Signaling Technology), and rabbit anti-vimentin (ab92547, Abcam).

Techniques: Stable Transfection, Expressing, Membrane, Staining, Control, Western Blot, Marker, Two Tailed Test