Fig. 1: Specific secreted proteins of V. cholerae impact the viability of cancer cells.
From: HapA protease targets PAR-1/2 to modulate ERK signalling and reduce cancer cell viability
A Schematic representation of the cancer cell viability assay. B Viability assay in MCF-7 cells co-cultured with the supernatant from each of the 6 V. cholerae mutant strains with reverted effect, the wild-type V. cholerae, or the E. coli strain at concentrations of 0, 0.5, 1 and 2.5% for 24 h. Live cells were fixed and stained using crystal violet, and cell quantity was measured at 590 nm. Experiments were performed in triplicate, results were normalized applying Log10 and data are presented as mean ± SEM. (*) p-value < 0.05; (**) p-value < 0.01; (***) p-values < 0.001; (n.s) p-value > 0.05. C Bacterial regulatory pathway relating the proteins with a negative impact on cell viability in the assay. D A autodegradation HapA protein (37 KDa) detection from the supernatant of wild-type V. cholerae, its isogenic ΔhapA mutant, the E. coli K12 strain MC1061 habouring the pBAD18 vector, and MC1061 expressing HapA from V. cholerae. E Coomassie blue staining of the gel served as a loading control for the immunoblot shown in (D). F Western blot analysis showing HapA expression levels in supernatants isolated from A1552 wild-type and its isogenic mutants (ΔhapA, ΔhapR, Δcrp, Δcya, ΔrpoS, ΔprtVΔhapA, Δvcc, ΔprtV, ΔtcpA, ΔwavB, and ΔtoxR).
