Fig. 1

RAGE LPA-RAGE axis mediates cellular proliferation, migration and invasion via PKB signaling pathway in lung cancer cells. a Levels of p-ERK/ERK, and p-PKB (Ser473)/PKB, in A549 cells treated with LPA for the indicated time points. b Levels of p-ERK/ERK, p-PKB (Ser473)/PKB and GAPDH in A549 cells treated with LPA (5 μM) for the indicated time points after shRNA mediated RAGE silencing. c Immunoblots showing levels of cyclinD1, c-myc, twist and GAPDH in A549 cells treated with LPA for the respective time points in hours. d Wound healing assay showing the migration of A549 cells stimulated with LPA in untreated, control shRNA and RAGE shRNA transfected cells. e Quantitative representation of cell proliferation via MTT assay comparing OD at 570 nm in control shRNA and RAGE shRNA transfected A549 cells in presence and absence of LPA (5 μM) for indicated time points. f Transwell invasion of A549 cells after RAGE silencing upon LPA treatment and its quantification. g Representative images and quantification of colonies of A549 cells transfected with control shRNA and RAGE shRNA and then grown in medium with or without LPA for 10 days. h F-actin polymerization in control and RAGE shRNA A549 cells cultured in medium with or without LPA for 6 h (green, F-actin; blue, DAPI). i Wound healing assay showing the migration of RAGE silenced A549 cells pre-treated with PKB inhibitor (LY294002) followed by no treatment (control), LPA and 5% FBS treatment. j Quantification of cell proliferation assay of control and RAGE silenced A549 with LPA post PKB inhibitor treatment (LY294002). k Trans-well invasion of RAGE silenced A549 cells pre-treated with PKB inhibitor (LY294002) followed by no treatment (control), LPA and 5% FBS treatment with their quantitative representations. l Immunohistochemical analysis and quantifications of % area DAB in sections of p-PKB (Ser473) in tumor sections of control shRNA and RAGE shRNA A549 cells intraperitoneally injected nude mice with exogenous LPA treatment. For all experiments, data are means ± SD, n = 3–7. **P ≤ 0.01, ***P ≤ 0.001