Fig. 5

a Characterization of KAIMRC1 cells on protein level. Western blot analysis of breast cancer cell lines, MCF-7, MDA MB-231 and normal breast cell line MCF-10A against AKT, p-AKT, ERK, p-ERK, p38 and p-p38. Interestingly, AKT was found to be constitutively active with or without starvation. b Human Phospho-Mitogen-activated Protein Kinase (MAPK) Antibody Array analysis of KAIMRC1 and MDA MB-231 cells. Phosphorylation of AKT was confirmed in KAIMRC1 cells as well as strong phosphorylation of GSK-3α/β was also observed whereas, MDA MB-231 cells showed active phosphorylation of ERK, GSK-3α/β and p38δ. c Bar graph representation of Human Phospho-MAPK antibody array. X-axis represents selected phospho-MAP Kinases and y-axis represents intensities of the dots visible in (b). All experiments were done in triplicates. d Effect of AKT/PI3K Pathway Inhibitors on KAIMRC1 Cells. Selective inhibitor of AKT signaling, API-2; Selective inhibitor of AKT/PKB pathway, 10-DEBC Hydrochloride and PI 3 Kinase inhibitor, LY294002 were used to inhibit AKT/PI3K pathway to study the constitutively active state of AKT in KAIMRC1 cells in comparison to MDA MB-231 and MCF-7 cell lines. CellTiter-Glo® assay was performed to assess cell viability after compound treatment. The cells were treated with compounds in complete media with and without serum 24 h prior to the assay. All the three cell lines showed no response to compound treatment suggesting that AKT pathway is not the only pathway involved in the carcinogenesis of KAIMRC1 cells. X-axis = Relative light unit (RLU) and y-axis = Logarithm of the drug molarity (Log[drug]M)