Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Cell viability ( A ) and cell proliferation ( B ) induced by nicotine in HBEpC and/or si-mRNA-α7-HBEpC viability. ( A ): For cell viability, 7500 cells/cm 2 are plated in T25 flask (total cell number 187,000) and treated with nicotine 1 × 10 −7 M, after 1 h cells are washed three times in PBS Ca 2+ and Mg 2+ free and then incubated in drug-free medium for additionally 48 h. Then, cells (detached or floating) are counted after staining with trypan blue dye. ( B ): For cell proliferation 7500 cells/cm 2 are plated in T25 flask and treated with nicotine every 48 h. Cells are detached and viable cells are counted every 12 h. Experiments are performed at least two times in triplicate. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Incubation, Staining, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Expression of Ki67 induced by nicotine in HBEpC and/or si-mRNA-α7-HBEpC. ( A ): ELISA experiments; ( B ): regression equation linearity, performed with Prism; ( C ): Western blotting, ( D ): densitometric analysis. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate. In the , raw data of Western blotting are reported.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: SA-β-Gal in HBEpC exposed continuously to nicotine. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques:

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Evaluation of intracellular Ca 2+ after exposure to nicotine for 48 h in HBEpC and/or si-mRNA-α7-HBEpC. ( A ): ELISA experiments; ( B ): regression equation linearity, performed with Prism. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Evaluation of intracellular ATP after exposure to nicotine for 48 h in HBEpC. ( A ): ELISA experiments; ( B ): regression equation linearity, performed with Prism. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Expression of EGF and p-EGFR after exposure to nicotine for 48 h in HBEpC. ( A ): ELISA experiments for EGFR; ( B ): regression equation linearity for EGFR, performed with Prism; ( C ): ELISA experiments for p-EGFR; ( D ): regression equation linearity for p-EGFR, performed with Prism. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Induction of p53 and phospho-p53 induced by nicotine in HBEpC. ( A ): ELISA assay. ( B ): Western blotting experiments, ( C ): densometric analysis. Experiments are performed at least two times in triplicate. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. In the , raw data of Western blotting are reported.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Induction of phospho-p38 and p38 by nicotine in HBEpC. ( A ): ELISA experiments; ( B ): regression equation linearity for phosphor-p38; ( C ): regression equation linearity for phosphor-p38performed with Prism. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: EMT induced by nicotine in HBEpC. ( A ): Western blotting; ( B ): densitometric analysis. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate. In the , raw data of Western blotting are reported.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Western Blot, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Cell migration to nicotine for 48 h in HBEpC and/or si-mRNA-α7-HBEpC. HeLa cells are positive control. ( A ) HeLa positive control. ( B ): Cell migration h in HBEpC and/or si-mRNA-α7-HBEpC. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Migration, Positive Control, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Induction of VEGF by nicotine in HBEpC and/or si-mRNA-α7-HBEpC. ( A ): ELISA experiments; ( B ) regression equation linearity for VEGFR. Statistical significance is analyzed with one-way AOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Anchorage-independent growth induced by nicotine in HBEpC. HeLa cells are positive control and NIH3T3 the negative. ( A ): Representative picture of HBEpC cloned on soft agar. ( B ): Cloned cells. Statistical significance is analyzed with one-way ANOVA with multiple-comparison and post hoc test with Bonferroni correction. Experiments are performed at least two times in triplicate.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Positive Control, Clone Assay, Comparison

Journal: Molecules

Article Title: Nicotine Changes Airway Epithelial Phenotype and May Increase the SARS-COV-2 Infection Severity

doi: 10.3390/molecules26010101

Figure Lengend Snippet: Effects induced by nicotine on different pathways in human airway epithelial cells (results obtained in this work and in literature) and comparison with effects caused by SARS-CoV-2, SARS-CoV, MERS-CoV and by non-tumorigenic virus infection on the same pathways.

Article Snippet: Human Bronchial Epithelial Cells (HBEpC) were obtained from Cell Applications Inc. ( www.cellapplications.com/product no. 502K-05a) and cultured in complete Bronchial/Tracheal Epithelial Cell Growth Medium ( www.cellapplications.com/product ) as described previously [ ]. si-mRNA-α7-HBEpC were obtained as described previously [ ].

Techniques: Comparison, Virus, Infection, Expressing, Concentration Assay, In Vitro, Activation Assay, Activity Assay, Knockdown, Control, Migration

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: Pharmaceuticals

Article Title: Corylin Ameliorates LPS-Induced Acute Lung Injury via Suppressing the MAPKs and IL-6/STAT3 Signaling Pathways

doi: 10.3390/ph14101046

Figure Lengend Snippet: Anti-inflammatory effect of corylin on LPS-induced ALI. The experimental results demonstrated that corylin attenuated the overproduction of IL-6 in LPS-activated human bronchial epithelial cells. In intratracheal LPS-induced ALI mice, corylin attenuated tissue damages, suppressed inflammatory cell infiltration, and decreased secretion of IL-6 and TNF-α in the BALF and serum; moreover, it further inhibited the expression of phosphorylation of mitogen-activated protein kinases (MAPKs), including the expression of p-JNK/JNK, p-ERK/ERK, p-p38/p38, and repressed the activation of signal transducer and activator of transcription 3 (STAT3) in lung. Taken together, our results are the first to demonstrate the anti-inflammatory effects of corylin on LPS-induced ALI and suggest corylin has significant potential as a novel therapeutic agent for ALI.

Article Snippet: A human bronchial epithelial cell line, HBEC3-KT cells, was purchased from American Type Culture Collection (No. CRL-4051, Manassas, VA, USA).

Techniques: Expressing, Phospho-proteomics, Activation Assay

Journal: Cell Death & Disease

Article Title: USP14 and UCHL5 synergistically deubiquitinate PKCα and translocate NF-κB to promote the progression of anaplastic thyroid cancer

doi: 10.1038/s41419-025-07890-9

Figure Lengend Snippet: A Expression levels of USP14 and UCHL5 in healthy thyroid and ATC tissues using the GEO database ( GSE33630 ). B IHC staining to evaluate the expression of USP14 and UCHL5 in normal thyroid, PTC, and ATC tissues. Scale bar, 100 μm. C Western blotting was used to detect USP14 and UCHL5 expression in one normal thyroid cell line (Nthy-ori3-1), two PTC cell lines (TPC-1, BCPAP), and six ATC cell lines (ARO, CAL62, KHM5M, 8505 C, 8305 C, and C643). D The qRT-PCR was used to detect the mRNA expression of USP14 and UCHL5 in normal thyroid, PTC and ATC cell lines. The experiment was repeated three times. E Correlation of USP14 and UCHL5 expression was analyzed by GEPIA2. F , G Kaplan-Meier plotter survival analysis illustrating the impact of USP14 and UCHL5 expression levels on the survival rates of thyroid cancer patients.

Article Snippet: The thyroid cell lines Nthy ori 3-1 (Fuheng Biotechnology), TPC-1 (Procell), BCPAP (Procell), 8505 C (Fuheng Biotechnology), C643 (DSMZ), CAL62 (DSMZ), ARO (Fenghui Biotechnology), and KHM5M (Procell) were cultured in RPMI-1640 medium with 10% FBS at 37 °C in 5% CO 2 .

Techniques: Expressing, Immunohistochemistry, Western Blot, Quantitative RT-PCR