Fig. 5.

Elevation of ER stress-related genes and intracellular Ca²⁺ level by FABP4 knockdown in the 3T3-L1 adipocytes. (A) RT-PCR analyses for ER stress-associated genes. At 48 h after siRNA transfection, the expression of several genes related to ER stress and/or UPR were analyzed by RT-PCR. Three independent samples transfected with siFabp4 or siControl were used in this evaluation. (B) Live cell calcium imaging. At 48 h after transfection of siFabp4 or siControl, the cells were stained with Fluo-8-AM (Green). Cell nuclei were counterstained with Hoechst33342 (Blue). Typical CLSM images of 3 independent experiments were shown. Scale bars represent 100 μm. (C) Quantification of intracellular Ca²⁺ level. Mean fluorescent intensity (FI) (average intensity of pixels per cell) for 67–73 adipocytes per condition was measured. Open and closed circles represent the mean FI values in each cell, and the black bars indicate the average values of the mean FI in each condition. ^#P < 0.0001 (Student’s t-test). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)