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. 2019 Mar 14;17(8):1670–1678. doi: 10.1111/pbi.13092

Figure 1.

Figure 1

Sl CYS8 inhibits papain‐like cysteine proteases (PLCPs) in the apoplast of N. benthamiana leaves. Apoplastic fluids from plants agroinfiltrated with an empty vector control (EV) or transiently expressing Sl CYS8 or its inactive mutant Q47 P S l CYS8 were isolated and incubated with MV201 or fluorophosphonate (FP)‐TAMRA probes to target (a) papain‐like cysteine proteases (PLCPs) and (b) Ser hydrolases (SHs), respectively. Labelled proteins were detected by in‐gel fluorescence scanning. Proteins were electrotransferred for immunodetection of Sl CYS8 in the apoplast. Arrows show signals with different fluorescence intensities in plants expressing Sl CYS8. As controls, apoplastic fluids were mixed and pre‐incubated with specific chemical inhibitors (E‐64 for PLCPs and DCI for SHs) before incubation with or without (Ctrl) the probes. Coomassie blue‐stained gels are shown as loading controls.