Journal: International Journal of Molecular Sciences

Article Title: Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum

doi: 10.3390/ijms23052488

Figure Lengend Snippet: M. mucogenicum morphologies. ( A ) Colony characteristics of M. mucogenicum in 7H10 media after 7 days. ( B ) Scanning electron microscopy images of the Mmuc -ATCC (ATCC 49650) strain and other clinical strains. ( C ) The purified GPLs and the acetone-precipitated pellet (APP) from total lipids extracted from Mmuc -ATCC (lane l), Mmuc -S (lane 2), and Mmuc -R (lane 3) were analyzed by thin-layer chromatography (TLC) using chloroform/methanol (9:1, v / v ) as the mobile phase. GPL: glycopeptidolipids.

Article Snippet: The Mmuc reference strain (ATCC49650) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Electron Microscopy, Purification, Thin Layer Chromatography

Journal: International Journal of Molecular Sciences

Article Title: Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum

doi: 10.3390/ijms23052488

Figure Lengend Snippet: Quantitative analyses of the necrosis and apoptosis of BMDMs infected with M. mucogenicum . Representative results of Annexin V–PI staining ( A ) and quantitative analysis ( B ). BMDMs were infected with Mmuc at an MOI of 10 for 24 h. After incubation, cells were collected, and FITC-Annexin V and PI were added. Samples were analyzed by flow cytometry. Values are mean ± SD of three samples (* p < 0.05, ** p < 0.01, *** p < 0.001 versus control group; ### p < 0.001 versus Mmuc-ATCC group).

Article Snippet: The Mmuc reference strain (ATCC49650) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Staining, Incubation, Flow Cytometry, Control

Journal: International Journal of Molecular Sciences

Article Title: Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum

doi: 10.3390/ijms23052488

Figure Lengend Snippet: M. mucogenicum -induced activation of MAPKs and NF-kB. ( A ) BMDMs were infected with the indicated Mmuc strains at an MOI of 10, and protein expression was detected at 30 min. Cell lysates were subjected to SDS–PAGE, and immunoblot analysis was performed using specific antibodies against phospho-p38 (p-p38), phospho-ERK1/2, phospho-JNK, and β-actin. ( B – D ) Each protein bands in A were scanned, and relative band intensities were normalized for the β-actin band. The column diagrams represent average relative band intensity with standard error from three independent experiments. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 versus control; # p ≤ 0.05, ## p ≤ 0.01 for Mmuc -ATCC versus Mmuc -S or Mmuc -ATCC versus Mmuc -R or Mmuc -S versus Mmuc -R). ns: nonsignificant.

Article Snippet: The Mmuc reference strain (ATCC49650) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activation Assay, Infection, Expressing, SDS Page, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum

doi: 10.3390/ijms23052488

Figure Lengend Snippet: Oxygen consumption rate of Mmuc -infected BMDMs. BMDMs were infected with Mmuc -ATCC, Mmuc -S, or Mmuc -R at an MOI of 10 for 24 h. ( A ) Mitochondrial respiration was determined while monitoring oxygen consumption rates (OCRs) with a Seahorse XFp analyzer using a Cell Mito stress test kit. The sequential injection of oligomycin (Oli, 1.0 μM), cyanide-4-[trifluoromethoxy] phenylhydrazone (FCCP, 2.0 μM), and rotenone/antimycin A (R/A, 0.5 μM) is indicated. ( B – G ) Representative nonmitochondrial oxygen consumption, basal respiration, maximum respiration, H + (proton) leakage, ATP production, and spare respiratory capacity values were determined using a Seahorse XF Cell Mito stress report generator. The data are normalized to the protein concentration. All data are presented as the mean ± SD ( n = 3). (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 versus control). ns: nonsignificant.

Article Snippet: The Mmuc reference strain (ATCC49650) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Injection, Protein Concentration, Control

Journal: International Journal of Molecular Sciences

Article Title: Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum

doi: 10.3390/ijms23052488

Figure Lengend Snippet: Extracellular acidification profiles and glycolytic parameters of Mmuc -infected BMDMs. BMDMs were infected with Mmuc -ATCC, Mmuc -S, or Mmuc -R at an MOI of 10 for 24 h. ( A ) Representative measurements of the extracellular acidification rates (ECARs) in Mmuc -infected BMDMs were acquired using the XF Glycolysis Stress Test kit. The sequential injection of glucose (Glu, 10 mM), oligomycin (Oli, 1.0 μM), and 2-deoxyglucose (2-DG, 50 mM) is indicated. ( B – F ) Representative glycolysis, glycolytic capacity, glycolytic reserve, nonglycolytic acidification, and glycolytic reserve as a percentage values were determined using the Seahorse XF Cell ECAR report generator. The data are normalized to the protein concentration. All data are presented as the mean ± SD (n = 3). (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001 versus control; # p ≤ 0.05 for Mmuc -ATCC versus Mmuc -R). ns: nonsignificant.

Article Snippet: The Mmuc reference strain (ATCC49650) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Injection, Protein Concentration, Control