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. 2024 May 7;27(6):109918. doi: 10.1016/j.isci.2024.109918

Figure 1.

Figure 1

Domain organization and bait two-hybrid analysis between human cDNA library and PfPHB1/PfPHB2

(A) Schematic representation of predicted domains of PfPHB (i) and PfPHB (ii). Stomatin-prohibitin flotillin-HflC/K (SPFH) superfamily bearing the common PHB domain is depicted.

(B) (i) X-gal indicator plate of bacterial two-hybrid experiment between human cDNA library and PfPHB1 and PfPHB2. Plates are labeled to represent genes cloned in pTRG/pBTnn. ESAT6pTRGnn/CFP10pBTnn is the positive control while ESAT6/empty pBTnn is the negative control. (ii) Protein parameters highlighting length, molecular weight (MW), isoelectric point (pI), number of cysteines, and grand average of hydropathicity (GRAVY) of the identified host-pathogen protein interactors. (iii) Liquid β-galactosidase assay for quantitative estimation of interaction strength between PfPHB2 and Hsp70A1A compared to that with positive and negative controls. Co-transformants ESAT6pTRGnn/CFP10pBTnn and ESAT6/empty pBTnn were taken as positive and negative control, respectively. Enzyme activity is expressed in terms of Miller units. The graph is the average of three independent assays, and standard deviation is represented by error bars. Statistical analysis was performed using unpaired t test and is shown with p values ≤0.0001∗∗∗∗.