Fig. 4
Microscale thermophoresis (MST) demonstrates a direct interaction between HIV-1 RT inhibitors and RED fluorescent-tagged TNF-α. For analysis, the change in thermophoresis is expressed as the change in the normalized fluorescence (ΔFnorm), which is defined as Fhot/Fcold (F-values correspond to average fluorescence values between defined areas marked by the red and blue cursors). Titration of the non-fluorescent ligand results in a gradual change in thermophoresis, which is plotted as ΔFnorm to yield a binding curve, which can be fitted to derive binding constants. A Experimental MST traces of efavirenz (KD = 24 ± 8 µM); B Experimental MST traces of delavirdine (KD = 39 ± 9 µM); C Experimental MST traces of nevirapine. Only the best MST traces (highest signal to noise ratio) are shown here. Values for all experiments conducted according to different experimental protocols are listed in Additional file 1: Table S4