Journal: Nature Communications

Article Title: PMI-controlled mannose metabolism and glycosylation determines tissue tolerance and virus fitness

doi: 10.1038/s41467-024-46415-4

Figure Lengend Snippet: a Growth curves of A549-wildtype (WT) and knockout ( PMI -/- ) cells supplemented with 0, 5, or 25 mM of mannose ( n = 4 biological repeats). b Bioenergetic profiles of H1N1-infected WT and PMI -/- cells supplemented with or without mannose (25 mM) treatment. Cells were pre-treated with mannose overnight before virus infection for another 12 h. Glycolytic rate of the cells were analyzed using a Seahorse XF analyzer by measuring the glycoPER kit ( n = 5 biological repeats). c Mitochondrial respiration of the cells was analyzed using a Seahorse XF analyzer by measuring the oxygen consumption rates (OCR, n = 5 biological repeats). d Either PMI overexpression (O/E) or supplement of PMI metabolite Fructose-6-phosphate (F6P) antagonize the suppression of low mitochondrial membrane potential (MMP) by mannose. A549 cells were pre-treated with the indicated treatment for overnight before H1N1 infection (MOI = 2). After 12 h, cells were subject to MMP analysis after JC-1 staining ( n = 3 biological repeats). e Dual PMI depletion and mannose treatment is unfavorable whereas supplement of F6P reverses the mitochondrial damage. A549-WT and PMI –/– cells were pre-treated with the indicated treatment, followed by H1N1 infection and MMP measurement after JC-1 staining ( n = 3 biological repeats). f MMP assay by TMRE Staining. hBTEC (left panel) and A549 cells (right panel) were pre-incubated with the indicated treatment for 12 h before H1N1 virus infection (MOI = 0.2). Cells were subject to TMRE (200 nM) staining for 20 min, and DAPI staining for normalization. The fluorescence intensities were detected by a plate reader, whereas the images were captured by GE IN Cell Analyzer 6500HS ( n = 3 biological repeats). g Knockout of PMI impair the modulatory activity of mannose against IL-1β production. The experiments were performed in WT and PMI –/– cells transfected with PMI plasmid and/or Poly(I:C) as indicated ( n = 3 biological repeats). All the results are shown as mean ± SD. One-way ANOVA with Dunnett’s post hoc test was used for ( a , d , e ). Unpaired and two-sided student’s T test was used for ( f , g ). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, and n.s. indicates non-significant.

Article Snippet: The Bronchial/Tracheal Epithelial Cells (hBTEc) were obtained from the ATCC company (ATCC PCS-300-010) and were cultured with ATCC complete growth media, consisting of ATCC Airway Epithelial Cell Basal Medium (ATCC PCS-300-030) supplemented with Bronchial Epithelial Cell Growth Kit (ATCC PCS-300-040).

Techniques: Knock-Out, Infection, Virus, Over Expression, Membrane, Staining, Mmp Assay, Incubation, Fluorescence, Activity Assay, Transfection, Plasmid Preparation

Journal: Viruses

Article Title: Dissecting Phenotype from Genotype with Clinical Isolates of SARS-CoV-2 First Wave Variants.

doi: 10.3390/v15030611

Figure Lengend Snippet: Figure 2. Evaluation of SARS-CoV-2 variants in primary epithelial cells and K18-hACE2 mouse model. (A) Normal human primary tracheal epithelial cells (NHTE) and normal human primary nasal epithelial cells (NHNE) were inoculated with a MOI = 1 and virus titer were assessed at 1 to 3 dpi with a plaque assay. Data are represented as mean ± standard error of the mean. Virus isolates within the A lineage, WA1/2020 (red), and A.1, UT21 (green), are represented by dashed lines. Virus isolates within the B.1 lineage, UT12 (black), UT23 (navy blue), UT27 (orange), and UT29 (violet), and B.1.577, UT5 (brown), are represented by solid lines. In B-D, female K18-hACE2 mice were intranasally challenged at 104 PFU with either SARS-CoV-2 WA1/2020 (n = 2) or one of six clinical isolates UT5, UT12, UT21, UT23, UT27, or UT29 (n = 3). Graphs of lung viral titers (B), weight and body temperature (C), or selected immune responses in lung (D) are presented.

Article Snippet: Normal human tracheal epithelial cells (NHTE) donated from a 42-year-old Caucasian male were purchased through Cell Applications Cat# 504-05a, Lot# 2559.

Techniques: Virus, Plaque Assay

Journal: Cell reports

Article Title: Interferon (IFN)-γ promotes monocyte-mediated lung injury during influenza infection

doi: 10.1016/j.celrep.2022.110456

Figure Lengend Snippet: Key Resources Table

Article Snippet: Primary Bronchial/Tracheal Epithelial Cells; Normal, Human , ATCC , Cat#PCS-300-01.

Techniques: Virus, Recombinant, cDNA Synthesis, SYBR Green Assay, Protein Extraction, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Software, Microscopy, Flow Cytometry, Fluorescence

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 1: ENDS aerosol exposure causes increased cell death in human lung epithelial cells The BEAS-2B cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 2: ENDS aerosol exposure causes increased cell death in mouse lung epithelial cells The MLE12 cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 3: One week of ENDS aerosol exposure causes decreased trans-epithelial resistance, dissociation of cells, and cell death Primary human bronchial epithelial cells were differentiated on transwell inserts into a pseudostratified epithelial layer. These cells were exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N- ), HEPA filtered air (Air), 100uM nicotine (Nicotine), and room temperature (Untreated) for 4 min. Trans- epithelial resistance was measured using a WPI EVOM TEER meter and STX3 electrode. Measurements were taken in triplicate and averaged. Background readings of empty wells as well as baseline measurements of each sample were used to calculate the change in resistance over the course of the treatment. Dissociated and dead cells were detected using trypan blue exclusion and counting on a hemocytometer. Treatments occurred once per day for 7 days. Experiments were performed 2 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol

Journal: Annals of Medicine

Article Title: Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma

doi: 10.1080/07853890.2021.1934725

Figure Lengend Snippet: Hsa_circ_0042823 was up-regulated in AMC-HN-8 and TU686 cells. (A) The expression of hsa_circ_0042823 in HBECs, AMC-HN-8 and TU686 cells was assessed through qRT-PCR. (B and C) The qRT-PCR was performed to detect hsa_circ_00428233 expression in the AMC-HN-8 and TU686 cells after hsa_circ_0042823 overexpression or knockdown. ** p < .01 compared with HBECs; ## p < .01 compared with Vector; $$ p < .01 compared with Si-NC.

Article Snippet: Human bronchial epithelial cells (HBECs) were purchased from ATCC (Cat: PCS-300-013, Manassas, VA).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Knockdown, Plasmid Preparation