Storage lipid studies in tuberculosis reveal that foam cell biogenesis is disease-specific
Fig 6
Effects of TNFR neutralizing antibodies on mTORC1 and caspase activity.
MDM were treated with antibodies against TNFR1 or TNFR2 prior to infection. After 24 h of infection, whole cell lysates and RNA were obtained. (A) Analysis of phosphorylation state of mTORC1 and abundance of pro-caspase 8 by western blot. p-mTOR, band reacting with an antibody specific for mTOR phosphorylated at Ser-2448; mTOR, band reacting with an antibody recognizing total mTOR protein. Western blot bands were quantified by using ImageJ software. Numbers below each band indicate the intensity ratio of the test band relative to β-actin (loading control). Full-length blots are presented in S8 Fig. (B) Quantification of the western blot data in panel A. The graph shows the effect of TNFR antibody treatment (treated vs untreated) on the ratios of p-mTOR/mTOR and procaspase 8 abundance. The average and standard deviation for three donors is shown. (C) Gene expression analysis. Methods are described in the legend to Fig 3. The graph shows the median of five donors, with each dot representing one donor. Statistical significance (*p < 0.05, **p < 0.01) was assessed by one-sample (panel B) and paired (panel C) student t-tests. The comparisons in the paired tests are as indicated; the comparison in the one-sample student t-test was between treated and untreated cells. [It is noted that the decrease of 50% in the ratio of p-mTOR/mTOR observed with the anti-TNFR1 antibody treatment did not reach statistical significance (p = 0.09)]. Mtb: M. tuberculosis, TNFR1 AB: TNFR1 neutralizing antibodies, TNFR2 AB: TNFR2 neutralizing antibodies.