Fig 1: IL-1ß production varies in response to different M. bovis strains. (A and B) Average IL-1B mRNA fold change (A) and average IL-1ß protein release (B) by bMDM infected with AF2122 (green squares) and G18 (blue triangles). Error bars denote the standard errors from five biological replicates. An asterisk denotes that the expression at that time point is statistically significantly different from the level of that transcript or protein in uninfected bMDM. Black circles denote time points where there is a statistically significant difference in IL-1ß mRNA or protein levels in response to the two different M. bovis strains. The statistical significance of M. bovis strain over the whole time course is indicated on each graph. (C) Example of a flow cytometry histogram illustrating the shift in FAM-FLICA staining, indicative of CASP1 activation, of bMDM infected with AF2122 (green line) and G18 (blue line) compared to uninfected bMDM (gray line). (D) Histogram illustrating the average mean fluorescence intensities (MFI) of uninfected bMDM (NC) and bMDM infected with M. bovis strains. Error bars denote the standard errors from four biological replicates. Two asterisks denote that there was a statistically significant difference in CASP1 activity compared to that of uninfected bMDM by GLM and subsequent Fisher's test (P < 0.001). (E) Western blot illustrating pro-IL-1ß protein levels in bMDM cell lysates 24 h after M. bovis infection. U denotes uninfected, and A and G denote bMDM infected with AF2122 and G18, respectively. bMDM 1 and 2 relate to bMDM isolated from different biological replicates. ß-Actin was included as a loading control. (F) Western blot illustrating pro-IL-1ß and mature IL-1ß protein present in bMDM supernatants collected 48 h after M. bovis infection. Numbers relate to bMDM isolated from different biological replicates.