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

Journal: Molecular Medicine

Article Title: The therapeutic potential of Apigenin in amyotrophic lateral sclerosis through ALDH1A2/Nrf2/ARE signaling

doi: 10.1186/s10020-024-00977-7

Figure Lengend Snippet: Effect of APG on viability and apoptosis in NSC34 neuronal cells. A CCK8 assay to evaluate the effects of different doses of APG on the viability of NSC34 cells in vitro. B Flow cytometry analysis of apoptosis in NSC34 cells harboring the SOD1*G93A mutation treated with APG. C Western blot analysis of apoptosis-related proteins (Bcl-2, caspase-3, Bax) in SOD1*G93A mutant NSC34 cells treated with APG. D Flow cytometry assessment of ROS levels in SOD1*G93A mutant NSC34 cells under APG treatment. E - H ELISA for quantification of OS markers (MDA, 4-HNE, SOD, ALDH) in SOD1*G93A mutant NSC34 cells treated with APG. I qRT-PCR analysis of ALDH1A2 mRNA levels, and (J) Western blot analysis of ALDH1A2 protein levels in APG-treated SOD1*G93A mutant NSC34 cells. * P < 0.05, ** P < 0.01, * P < 0.001

Article Snippet: 293FT and NSC34 cell lines from Procell (Wuhan, China) were cultured in DMEM containing 10% fetal bovine serum (FBS) at 37 °C.

Techniques: CCK-8 Assay, In Vitro, Flow Cytometry, Mutagenesis, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Molecular Medicine

Article Title: The therapeutic potential of Apigenin in amyotrophic lateral sclerosis through ALDH1A2/Nrf2/ARE signaling

doi: 10.1186/s10020-024-00977-7

Figure Lengend Snippet: Effects of ALDH1A2 knockdown and APG treatment on ALS model and OS markers. A - B Validation and selection of the most effective sh-ALDH1A2 to suppress ALDH1A2 expression by qRT-PCR and Western blot analysis. C Flow cytometry to evaluate apoptosis in NSC34 cells treated with APG and sh-ALDH1A2. D Western blot to evaluate the levels of apoptosis-related proteins (Bcl-2, caspase-3, Bax) in NSC34 cells treated with both APG and sh-ALDH1A2. E Flow cytometric analysis of ROS levels in treated NSC34 cells. F - I ELISA for quantification of OS markers (MDA, 4-HNE, SOD, ALDH) in NSC34 cells treated with APG and sh-ALDH1A2. J Western blot analysis of ALDH1A2 protein levels. K - L qRT-PCR and Western blot analysis of Nrf2, HO-1, NQO1 mRNA and protein levels in NSC34 cells treated with APG and sh-ALDH1A2. * P < 0.05, ** P < 0.01, * P < 0.001

Article Snippet: 293FT and NSC34 cell lines from Procell (Wuhan, China) were cultured in DMEM containing 10% fetal bovine serum (FBS) at 37 °C.

Techniques: Knockdown, Biomarker Discovery, Selection, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay