hqno  (TargetMol)

Journal: bioRxiv

Article Title: An effective response to respiratory inhibition by a Pseudomonas aeruginosa excreted quinoline promotes Staphylococcus aureus fitness and survival in co-culture

doi: 10.1101/2025.03.12.642861

Figure Lengend Snippet: Panels A and B. The wild type (WT; JMB1100) strain containing the pOS_ nrd_gfp (panel A) or pOS_ srrA_gfp (panel B) transcriptional reporters were co-cultured with different concentrations of HQNO before gfp expression was quantified using fluorimetry. Panels C and D. The WT and Δ srrAB (JMB1467) strains containing the pOS_ nrdD_gfp (panel C) or pOS_ srrA_gfp (Panel D) transcriptional reporter trains were cultured in TSB-Cm with and without 5 μg mL -1 HQNO before gfp expression was quantified using fluorimetry. Each individual data bar corresponds to the average of the biological replicates, and standard deviations are shown, but the data points sometimes obscure them. Student’s t-tests were performed and * denoting a p-value ≤ 0.05. BDL denotes below the detectable limit of the fluorimeter.

Article Snippet: Cells were treated with 5 μg mL -1 HQNO (Selleck Chemicals) prepared as a 5 mg mL -1 stock in DMSO or vehicle control.

Techniques: Cell Culture, Expressing

Journal: bioRxiv

Article Title: An effective response to respiratory inhibition by a Pseudomonas aeruginosa excreted quinoline promotes Staphylococcus aureus fitness and survival in co-culture

doi: 10.1101/2025.03.12.642861

Figure Lengend Snippet: Panel A. Oxygen consumption rates were quantified in the wild type (WT; JMB1100) and Δ cydA qoxB::tet (JMB 8988) strains after culture in TSB media with the presence or absence of 5 μg mL -1 HQNO. Panel B. Membrane potential was qualitatively assessed using 1 mM of the fluorescent dye DIOC 2 in the WT and Δ cydA qoxB : :tet strains after culture in TSB media with the presence or absence of 5 μg mL -1 HQNO. Panels C and D. The WT, Δ srrAB (JMB1467), Δ cydA qoxB : :tet , and Δ srrAB Δ cydA qoxB : :tet (JMB 8989) strains containing pOS_ nrdD_gfp or pOS_ srrA_gfp after culture in TSB media in the presence and absence of 5 μg mL -1 HQNO before gfp expression was quantified using fluorimetry. Each individual data bar corresponds to the average of the biological replicates, and standard deviations are shown, but the data points sometimes obscure them. Student’s t-tests were performed, and * denoting a p-value ≤ 0.05, and ns denotes no significant difference. BDL denotes below the detectable limit.

Article Snippet: Cells were treated with 5 μg mL -1 HQNO (Selleck Chemicals) prepared as a 5 mg mL -1 stock in DMSO or vehicle control.

Techniques: Membrane, Expressing

Journal: bioRxiv

Article Title: An effective response to respiratory inhibition by a Pseudomonas aeruginosa excreted quinoline promotes Staphylococcus aureus fitness and survival in co-culture

doi: 10.1101/2025.03.12.642861

Figure Lengend Snippet: Panel A. The relative abundances of NAD + and NADH in the wild type (WT; JMB1100) strain after co-culture in the presence and absence of 5 μg mL -1 HQNO. Panel B. The expression of gfp was monitored in the WT, Δ srrAB (JMB1467), Δ rex::kan (JMB 13604), and Δ srrAB Δ rex::kan (JMB 13626) strains containing pOS_ srrA_gfp after culture in the presence and absence of 5 μg mL -1 HQNO. The response to HQNO requires both SrrAB and Rex regulators. Each individual data bar corresponds to the average of the biological triplicates, and standard deviations are shown, but the data points sometimes obscure them. Student’s t-tests were performed, and * denoting a p-value ≤ 0.05, and ns denotes no significant difference. BDL denotes below the detectable limit.

Article Snippet: Cells were treated with 5 μg mL -1 HQNO (Selleck Chemicals) prepared as a 5 mg mL -1 stock in DMSO or vehicle control.

Techniques: Co-Culture Assay, Expressing

Journal: Nature Communications

Article Title: A genome-wide CRISPR-Cas9 knockout screen identifies FSP1 as the warfarin-resistant vitamin K reductase

doi: 10.1038/s41467-023-36446-8

Figure Lengend Snippet: a Scheme of the FSP1-CoQ 10 -NADH and DHODH-CoQ 10 -DHO pathways for ferroptosis suppression. CoQ 10 can be reduced to CoQ 10 H 2 by either FSP1 or DHODH using the reducing equivalent from NADH or DHO, respectively. Reduced CoQ 10 traps lipophilic radicals to suppress ferroptosis. NADH reduced nicotinamide adenine dinucleotide, NAD+ oxidized nicotinamide adenine dinucleotide, DHO dihydroorotate, OA orotic acid, PLOOH phospholipid hydroperoxides, PLOO· phospholipid peroxyl radical. b Cell-based VKR activity of FSP1 and DHODH. FSP1 and DHODH were transiently expressed in TKO reporter cells for 24 h for carboxylation activity assay. Wild-type FSP1 activity was normalized to 100%. c Effect of FSP1 inhibitors on VKD carboxylation. Increasing concentrations of FSP1 inhibitors, HQNO or iFSP1, with 11 µM vitamin K were incubated with DKO reporter cells for 24 h for VKD carboxylation activity assay. Carboxylation activity of cells incubated with 11 µM vitamin K without inhibitor was normalized as 100%. d Effect DHODH inhibitors on VKD carboxylation. Increasing concentrations of DHODH inhibitors, BRQ and LFM, with 11 µM vitamin K were incubated with DKO reporter cells for 24 h for VKD carboxylation activity assay. Carboxylation activity of cells incubated with 11 µM vitamin K without inhibitor was normalized as 100%. e Lineweaver–Burk plot of VKR activity in DKO cells as a function of vitamin K concentration in the absence and presence of HQNO. DKO reporter cells were cultured with increasing concentrations of vitamin K containing 0, 0.625, 1.25, 2.5, 5.0, and 10 μM HQNO for the activity assay. Data were plotted using Graphpad software. Data are presented as mean ± SD of three independent experiments ( n = 3) in Fig. 5b–d.

Article Snippet: HQNO (2- n -heptyl-4-hydroxyquinoline- N -oxide, #15159), iFSP1 (#29483), Brequinar (BRQ, #36183) and Leflunomide (LFN, #14860) were sourced from Cayman Chemical.

Techniques: Activity Assay, Incubation, Concentration Assay, Cell Culture, Software