Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol before being subcultured into fresh media at an optical density (OD 600 ) of 0.05. Bacterial growth was monitored by measuring the OD 600 every 24-hours for 7 days.

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques:

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol before being subcultured into fresh media at an optical density (OD 600 ) of 0.05. Bacterial growth was monitored by measuring the OD 600 every 24-hours for 7 days and plotted on a log transformed scale (A). The same strains were examined for growth in sodium dodecyl-sulfate (SDS) by subculturing exponential phase bacteria into 7H9 supplemented with 10% OADC and 0.2% tyloxapol with and without 0.2% SDS at an OD600 of 0.8. Twenty-four hours later, these cultures were serially diluted and plated onto 7H11 agar plates supplemented with 10% OADC in technical triplicate. Images are representative of three biological replicates each plated in technical triplicate (B). Colony forming units were quantified from agars plates following incubation at 32°C with 5% CO 2 for one week (C). Statistical significance was determined using a one-way ANOVA followed by a Dunnett’s test for multiple comparison relative to WT, with p-values ≤ 0.05 considered significant (C only).

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques: Transformation Assay, Subculturing Assay, Bacteria, Incubation, Comparison

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol and used to infect triplicate wells of BMDMs, at an MOI of 0.2, for 2 hours. At 2, 48, and 96 hours post infection (hpi), macrophage monolayers were lysed and plated onto 7H11 agar supplemented with 10% OADC in technical triplicate. One week after plating, colony forming units were enumerated for each timepoint (A). Bacterial numbers present at 2hpi were plotted separately (B) to better visualize differences in bacterial counts. Uninfected wells were included as a control for across well contamination (not shown, no contamination observed) and the Δ esxBA strain as a control for attenuated growth. Statistical significance was determined using a Kruskal-Wallis test followed by a Wilcoxon Rank Sum test for pairwise comparison relative to WT, with p-values ≤ 0.05 considered significant (*** p<0.001).

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques: Infection, Control, Comparison