Fig. 8.

Western blot analysis of TfR2 and TMPRSS6 protein levels in IL-6 neutralized and LPS or LTA treated BV2 cells. BV2 cells were fractionated upon collection using the Subcellular Protein Fractionation Kit. Protein contents of the fractions were measured and the same amount of protein from each sample was loaded onto 10% polyacrylamide gels and were separated by electrophoresis, then were transferred by electroblotting onto nitrocellulose membranes. The membranes were probed with rabbit polyclonal antibodies produced against TfR2 and TMPRSS6 according to the manufacturer’s protocols. β-actin was used as housekeeping control. a Western blot analyses of TfR2 and TMPRSS6. b Optical density analyses of TfR2 and TMPRSS6. Optical densities were calculated by ImageJ software and expressed as percentage of target gene/β-actin abundance. The columns represent mean values and error bars represent standard deviation (SD) of three independent experiments (n = 3). Ctrl untreated control, NCtrl control + neutralizing IL-6 antibody, NLPS neutralizing IL-6 antibody + LPS, NLTA neutralizing antibody + LTA. The asterisk marks p < 0.05 compared to Ctrl. The cross marks p < 0.05 compared to NCtrl. The double cross indicates significant difference, p < 0.05 between LPS and NLPS or LTA and NLTA. Data was analysed by two-way ANOVA followed by Tukey’s HSD post hoc test