Fig. 6
From: Increased yields and biological potency of knob-into-hole-based soluble MHC class II molecules
Use of a c-jun/c-fos zipper in KIH-based pMHCII is incompatible with expression. a, b Cartoons displaying the structure of the two types of KIH constructs tested here. c (left), Expression of eGFP in CHO-S cell lines transduced with lentiviruses encoding the constructs depicted in A (top) and B (bottom), indicating adequate construct transcription and translation. c (right), FPLC elution profiles of pMHC class II from Strep-tactin columns loaded with supernatants from CHO cells expressing the constructs in A (top) or B (bottom). d Effects of the KIH on the SDS stability of a representative pMHCII heterodimer (out of at least 10 different KIH-based pMHCs), in the absence of Cys-trapping. Data correspond to c-jun/c-fos-based BDC2.5mi/IAg7 (‘conv’, left lane) and a KIH-based BDC2.5mi/IAg7 (right lane). βME, beta-mercaptoethanol. e Representative pMHCII tetramer/eGFP (TCR) FACS dot plots for BDC2.5-TCR-transgenic CD4+ T-cells stained with c-jun/c-fos- (‘conv’, left) or KIH-based BDC2.5mi/IAg7 tetramers (right). The staining patterns shown are representative of at least two independent experiments
