Metabolite composition differences and lipid droplet dynamics analysis in siRNA transfected 3T3-L1 differentiated cell

Abstract

To understand the connection between the function of lipid droplet controlling genes and phenotype in adipocytes, we used label free imaging technique based on CARS(Coherent anti-Stokes Raman spectroscopy), and combined ToF-SIMS(Time-of-flight secondary ion mass spectrometry) to obtain chemical information for further explanation. 3T3-L1 cells were transfected with four different kinds of siRNAs during differentiation. Abcd2, adrp, kif3a, plin genes were knockdowned temporarily and the effects on lipid droplet formation and dynamics were imaged with CARS. Metabolites were extracted from images samples were analyzed with ToF-SIMS to obtain mass spectrum. Data of four samples and control were compared with multivariate analysis method. siADRP sample showed the most extreme changes in phenotype, and siKIF3A sample changed the most in dynamics compared to control sample.