Fig. 1

Butyrate treatment decreases membrane cholesterol, increases fluidity and disrupts lipid rafts in RAW264.7 cells. RAW 264.7 cells were treated with 5 mM or 10 mM or without sodium butyrate for 18 h. The cells were washed and membrane was prepared. The membrane cholesterol was estimated by amplex red cholesterol assay kit (invitrogen) and expressed as µg of membrane cholesterol/ mg protein A, the percent reduction of membrane cholesterol (Inset). The cells treated with/without butyrate were stained with Fillipin and observed under fluorescence microscope B and measured in flowcytometry C, D. After 18 h of butyrate treatment the cells were further treated with or without chol-lipo for another 18 h. The membrane anisotropy was measured by using Laudran probe and expressed as r E. The cells were stained with either CTX-B-FITC or anti-CD71 antibody and counterstained with Hoechst 33342. The cells were imaged in Zeiss confocal microscope F. The quantitative analysis of total fluorescence of CTX-B G, and anti-CD71 antibody H, as measured by ImageJ. The corrected total cell fluorescence is calculated as Corrected total fluorescence (CTCF) = (Integrated density)—(Area of selected cells x mean fluorescence of background). Each experiment were set in triplicate and the data of three experiments are plotted as Mean ± SEM. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001