Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) CD11c + cells are increased in the lungs of atopic mice. Flow cytometry of lung cell suspension showing frequency (left) and cell number (right) at day 3 post inoculation (PI) high dose SeV ( B ) Adoptive transfer of CD11c + cells isolated from NA or atopic mice into naïve mice 24 h before inoculation with high dose SeV delays but does not prevent mortality; n=4 per group. ( C ) Transcriptomic (RNAseq) comparison between FACS isolated lung CD11c + cells from atopic and NA mice identifies several disparately expressed gene products, including Nrg1 (n=4 per group). Selected gene products shown. ( D ) NRG1 is markedly increased in atopic mouse lung (“tissue”), BAL, and supernatant from 1 x10 6 atopic lung CD11c + cells (“CD11c sup”) cultured for 24 h. ( E ) The scRNA (10x Platform) tSNE coordinates show CD14 + monocytes and dendritic cell (“DC”) sub-populations expressing NRG1 in peripheral blood cells of healthy human donors.*p<0.05, **p<0.01, ****p<0.0001

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Flow Cytometry, Suspension, Adoptive Transfer Assay, Isolation, Comparison, Cell Culture, Expressing

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) NRG1(1ng to 1000ng) i.n. (in 30µL) given daily to naïve mice for 5d before inoculation with high dose SeV reduces viral mortality; n=4 per group (1ng, 10ng 1000ng & PBS), n=8 per group (100 ng and 500 ng). ( B ) Ratio of EBD in the BAL to that in the lung shows reduced EBD in NRG1 treated mice on day 8 PI SeV. n≥8 per group, median ± IQR shown, Mann-Whitney U test.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) Adding NRG1 to hBEC inoculated with rgRSV (left) and mTEC inoculated with GFP-SeV (right) reduces spread of infection. Representative images shown. ( B ) Quantification of (A) for rgRSV and hBEC and ( C ) for GFP-SeV and mTEC. GFP positive cells quantified by ImageJ. Representative images from ≥3 separate experiments. *p<0.05, **p<0.01. ( D ) Transcriptomic analysis of hBEC cultures treated with NRG1 (100 ng) on the basolateral side of the Transwell for 5 days and inoculated with RSV (4000 pfu). RNA was isolated 48 h PI RSV and qRT-PCR performed using a custom Prime PCR array plate: (i) Transcripts in which NRG1 treatment reduced gene expression from that seen in RSV infected cells. (ii) Transcripts with low level expression that show small but significant change in expression relative to naïve control with NRG1 alone or genes significantly increased with RSV but whose expression levels were not affected by NRG1. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n=3.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Infection, Isolation, Quantitative RT-PCR, Gene Expression, Expressing, Control

Journal: eLife

Article Title: NF-κB oscillations translate into functionally related patterns of gene expression

doi: 10.7554/elife.09100

Figure Lengend Snippet: Figure 1. Periodic forcing turns damped heterogeneous oscillation into synchronous sustained oscillations. (A) The activity of NF-kB is regulated through different negative feedbacks provided by the inhibitors IkB and A20. The scheme at the bottom represents a generic forcing with periodically alternating TNF-a doses D1 and D2 of duration T2+T1 = Tf; Tf is the period of the forcing. (B, C) Oscillations observed in three GFP-p65 cells obtained by computing the nuclear to the cytoplasmic GFP intensity (NCI) for constant flow of 10 ng/ml TNF-a (B) and upon alternating doses D1=10 ng/ml TNF-a, D2=0 ng/ml and T1=T2 =45 min (C). Each colour corresponds to a single cell trace. Oscillatory patterns can be effectively visualised using the phase f of the oscillation, which is 2p in the maxima of the oscillatory peaks (yellow) and p in the local minima (green) in the colour phase-plot of f(t) below the panels. Scale-bar for f is on the right. (D) Time lapse images of cells under constant stimulation displaying the characteristic heterogeneous nuclear-to-cytoplasmic translocations. (E) Phase plot drawn for 50 cells, of 105 analysed, showing the asynchrony of the oscillations except for the first peak. (F) Distribution of the experimentally computed period of the oscillations Texp, measured as the time between two consecutive oscillatory peaks. The distribution has a maximum at T0 =90 min, which corresponds to the natural period. (G) Quantification of the height for each peak. (H) Time lapse images of the cells under periodic stimulation, showing synchronous NF-kB Figure 1 continued on next page

Article Snippet: For WB, we used an anti p65 Rab (dil 1:1000, #sc-372 C20, Santa Cruz; Figure 1—figure supplement 10).

Techniques: Activity Assay

Journal: eLife

Article Title: NF-κB oscillations translate into functionally related patterns of gene expression

doi: 10.7554/elife.09100

Figure Lengend Snippet: Figure 5. Synchronous NF-kB oscillations lead to population-level coordinated transcription. (A) NCI plot of single cell oscillations (green lines) and population average (black lines) for cells stimulated with D1=10 ng/ml TNF-a, D2=0 ng/ml, T1=30 min and T2=150 min. The open circles represent the fitting obtained using our minimal mathematical model. (B) The mathematical model predicts waves of transcription (orange plot, right) coordinated with the stimulus (red plot, top) and p65 oscillations (green plot, left). (C, D) q-PCR time course of nascent and mature mRNAs for the prototypical early and late genes IkBa and Ccl5, respectively. (E, F) Transcription profiles for mature IkBa (red) and Ccl5 (blue) RNAs (dots) can be accurately fitted (lines) by our minimal mechanistic mathematical model. The fittings were performed by keeping common the parameters regulating the external signal (PS) and the dynamics (PNF-kB) in (A), (E) and (F) but using different gene expression parameters PG for (E) and (F). Figure supplements 1 to 2 are provided. DOI: 10.7554/eLife.09100.034 The following figure supplements are available for figure 5:

Article Snippet: For WB, we used an anti p65 Rab (dil 1:1000, #sc-372 C20, Santa Cruz; Figure 1—figure supplement 10).

Techniques: Gene Expression