Fig. 5.

Roles of intracellular signaling pathways and Mesothelin in glucose and insulin-invoked CA125 metabolism. a The computational simulation of Western blotting performed using the automatic machine “Wes”. Shown are the intracellular CA125 and Mesothelin protein levels after administration of saline, insulin and glucose plus insulin in ovarian cancer cells (OVCAR-3) with or without PI3K-Akt and MEK-ERK pathway blockades (for details, see Additional file 2: Figure S2). b The relative intracellular Mucin16 and MSLN mRNA levels after administration of saline, insulin and glucose plus insulin with or without PI3K-Akt and MEK-ERK blockades (note: Mucin 16/ACTB and MSLN/ACTB mRNA ratios were set to 1 for the saline controls). c Knockdown effects of MSLN-specific or NC siRNAs on target protein expression. d The time-dependent extracellular and intracellular CA125 levels after MSLN knockdown. e The time-dependent re-absorption of Alexa Fluor 488-labeled CA125 after MSLN knockdown. Only the cells treated with NC-siRNA and supplemented with insulin showed significantly increased re-absorption of CA125 protein. *Statistical significance (two-sided Student’s t test)