35G8 destabilizes PDI. (A) Thermal shifts observed for recombinant PDI (0.3 mg/ml) with various concentrations of 35G8. DMSO was used as a control. (B) Apparent melting temperatures (Tm) and change in melting temperature derived from ThermoFluor assay (C) Protein expression of PDI, GRP78, GSTO1, and actin (loading control) in the absence or presence of 35G8 at varying temperatures in the cellular thermal shift assay (D) Western blot analysis of DARTS assay with PDI, GRP78, and GSTO1 subjected to 100 μM PACMA31 (P), 100 μM 35G8 (G), or DMSO (−). Samples were subjected to varying concentrations of pronase. Data are means from three independent experiments.