Journal: bioRxiv

Article Title: Macrophage-intrinsic MDA5-IRF5 axis drives HIV-1 icRNA-induced inflammatory responses

doi: 10.1101/2024.09.06.611547

Figure Lengend Snippet: THP-1/PMA macrophages were infected with Lailienv GFP/G (MOI 2) in the presence or absence of efaverinz (EFV, 1 µM), raltegravir (RAL, 30 µM), spironolactone (Spiro, 100 nM), and KPT335 (KPT, 1 µM). Cells and culture supernatants harvested at 3 dpi for (A) flow cytometry analysis to measure infection levels (¾GFP+) and (B) ELISA to measure IP-10 secretion. Infection levels (C) and IP-10 secretion (D) in THP-1/PMA macrophages infected with either WT (LailienvGFP/G) or HIV-1/M10 were determined at 3 dpi by flow cytometry and ELISA, respectively. (E) MAVS expression in THP1 cells transduced with shCTRL or shMAVS lentivectors was determined by western blot analy­ sis. (F, G) LaiiienvGFP/G infected THP-1/PMA macrophages and cell supernatants were harvested 3 dpi for flow cytometry analysis (F) and ELISA (G) to measure infection establishment (¾GFP+) and IP-10 secretion. Data is displayed as the means ± SEM with each dot representing an independent experiment. Statistical significance assessed via 1-way ANOVA with Dunnet’s multiple comparisons test (A-B), unpaired t-test (C-O, F-G). *: p < 0.05; **: p <0.01, ***: p < 0.001, ****: p < 0.0001, ns = not significant.

Article Snippet: In some experiments, THP-1/PMA macrophages were pre-treated (for 20 minutes) with efavirenz (1 μM, NIH AIDS Reagent Program), raltegravir (30 μM, Selleck Chemical #50-615-1), spironolactone (100 nM, Selleck Chemical, # S4054), and KPT 330 (1 μM, Selleck Chemical # 50-136-5156).

Techniques: Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Transduction, Western Blot

Journal: bioRxiv

Article Title: FrlP, an ABC type I importer component of Bacillus subtilis : regulation and impact in bacterial fitness

doi: 10.1101/2025.07.11.664333

Figure Lengend Snippet: (A) Growth kinetic parameters of B. subtilis WT and mutants Δ msmX (IQB495), Δ msmX ΔS1254 (ISN51), Δ msmX frlPstop (ISN72) and P spank(hy) frlP (ISN15) were measured in CSK minimal medium supplemented with 0.1 % (wt/vol) arabinotriose as sole carbon and energy source and are shown as doubling time (min). Values above 500 min were considered no growth ( , ). The results represent at least three independent experiments and error bars represent standard deviation of the mean. (B) Representation of relative expression of frlP in the respective strains compared to the WT in CSK minimal medium supplemented with 0.1 % (wt/vol) arabinose. Primers ARA925 and ARA926 were used for frlP and fold-change was normalized using primers ARA583 and ARA584 for the 16S gene. Error bars represent standard deviation of the mean from C t values of at least three independent assays. (C) Western blot analysis of FrlP (c.a. 41.4 kDa) accumulation in total cell extracts of different B. subtilis strains. P spank(hy) - frlP (ISN15) and Δ frlP (IQB618) were used as positive and negative controls, respectively. NZYColour Protein Marker II (NZYTech) was applied as standard, and it is partially represented (MWM). The uncropped image of this bolt is presented in Fig. S3. Statistical significance of doubling time and fold-change of different strains compared to the respective controls is indicated (*p<0.05; **p<0.01; ***p<0.001).

Article Snippet: 20 μg of total protein from each extract were loaded in a 12.5 % SDS-PAGE and run at constant electrical current (80 mA) for 50 min using NZYColour Protein Marker II (NZYTech) as standard.

Techniques: Standard Deviation, Expressing, Western Blot, Marker