Journal: Allergy, asthma & immunology research

Article Title: Serum Amyloid A1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients.

doi: 10.4168/aair.2022.14.1.40

Figure Lengend Snippet: Fig. 1. The production of SAA1 from AECs and the effects of SAA1 on proinflammatory cytokine production. SAA1 proteins were produced from AECs (SAECs and A549, respectively) in response to various stimuli (A, B), and in the coculture with PBNs, PBEs (C, D). SAA1-induced release of IL-6, IL-8 and S100A9 from AECs (SAEC and A549, respectively) (E-K). Data are presented as means ± standard error of mean (n = 6). P values were calculated by using one-way analysis of variance with Bonferroni's post hoc test. SAA1 enhanced the expression of p-ERK and p-p38 MAPK (L) in AECs (SAECs and A549, respectively). IL, interleukin; PBEs, peripheral blood eosinophils; PBNs, peripheral blood neutrophils; dex, dexamethasone; S100A9, S100 calcium-binding protein A9; p-ERK, phospho-extracellular signal-regulated kinase; ERK, extracellular signal-regulated kinase; p-p38 MAPK, phospho-p38 mitogen-activated protein kinase; MAPK, mitogen-activated protein kinase; AECs, airway epithelial cells; SAEC, small airway epithelial cells. *P < 0.050, †P < 0.010, ‡P < 0.001, ns = not significant.

Article Snippet: Epithelial cell culture and stimulation Two kinds of AECs were used in this study, primary small airway epithelial cells (SAECs) and human AEC line (A549), both of which were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Produced, Expressing, Binding Assay

Journal: Nucleic acids research

Article Title: Sequestration of ribosomal subunits as inactive 80S by targeting eIF6 limits mitotic exit and cancer progression.

doi: 10.1093/nar/gkae1272

Figure Lengend Snippet: Figure 9. Ov ere xpression of eIF6 observ ed primarily in high-grade in v asiv e bladder and breast cancers. ( A and B ) Western blot probed for eIF6 in human bladder cancer cell lines and healthy (normal) bladder epithelial cells. β-Tubulin used as loading control. Blots shown in panel (A) were quantitated and eIF6 le v els w ere normaliz ed to loading control and plotted (panel B) as FC o v er eIF6 le v els in normal bladder epithelial cells. Values indicate standard error of the mean from four independent experiments and significant differences for T24 ( P = 0.0065), UMUC3 ( P = 0.0132) and HT1197 ( P = 0.0017) determined by an unpaired t wo-t ailed t -test. ( C ) Images represent eIF6 expression in patient-derived benign and high-grade tumors by immunohistochemistry using anti-eIF6 antibody. Enlarged inset shows the presence of eIF6 in nucleoli and cytoplasm in high-grade cancers. ( D ) Images shown in panel (C) were quantitated and eIF6 expression in patient-matched benign tissues relative to high-grade cancers (six patients) were plotted. Significant differences were determined using an unpaired t wo-t ailed t -test ( P = 0.0063). ( E ) Plot shows eIF6 expression in unmatched benign tissues, high-grade and low-grade bladder cancers derived from patients. ( F and G ) Western blot represents high levels of eIF6 in high-grade invasive human triple negative breast cancer cell line. β-Tubulin used as loading control. Blots shown in panel (F) were quantitated and eIF6 levels were normalized to loading control and plotted (G) as FC o v er eIF6 le v els in normal (healthy) HME-1 cells. Values indicate standard error of the mean from four independent experiments and significant differences for MDA-MB-231 ( P = 0.001) determined by an unpaired t wo-t ailed t -test.

Article Snippet: The human bladder cancer cell lines HT1376, HT1197, UM-UC-3 and TCCSUP were maintained in EMEM media with 10% FBS and Penn / Strep, T-24 cell lines were maintained in McCoy’s 5A medium with 10% FBS and Penn / Strep and primary bladder epithelial (A / T / N): normal human (BdEC) cells were maintained in bladder epithelial cell basal medium (ATCC) supplemented with bladder epithelial g c i g b D W C [ a p 1 fl d t ( m a p c w m ( b l o β S 0 p B S C C e d s i S B m B s a ( S B a p a m t w s i i H 7 w f v D ow nloaded from https://academ ic.oup.com /nar/advance-article/doi/10.1093/nar/gkae1272/7933602 by guest on 30 D ecem ber 2024 rowth factors.

Techniques: Western Blot, Control, Expressing, Derivative Assay, Immunohistochemistry

Journal: Allergy

Article Title: Human Pulmonary Neuroendocrine Cells Respond to House Dust Mite Extract With PAR-1 Dependent Release of CGRP.

doi: 10.1111/all.16416

Figure Lengend Snippet: FIGURE 1 | Characterization of iPNEC and ePNEC cultures. (A) Quantitative RT-PCR showing expression of characteristic PNEC markers in human cultured iPSC and -HBEC-derived iPNEC (□ female, 22; ■ male, 32) and ePNEC (● male, 52; ▲ male, 56; ○ female, 55) and control HBEC at day 60 in ALI culture, respectively. (B) Mean fluorescent intensity (MFI) of CHGA+/Hoechst+ cells (left) and SYP+/Hoechst+ cells [34] at day 60. Data is representative of four different culture plate wells per group for one biological donor for each cell type (ePNEC male, 52; iPNEC male, 32). (C) Representative IF images show SYP+ cells and nuclei are counterstained with Hoechst. 20× magnification and scale bars at 100 μm. (D) Single- cell RNA sequencing identifies two ePNEC clusters in 60-day-old ePNEC differentiated cells. Each dot represents one well and data shown for mean ± SEM. Mann–Whitney test performed for (A, B); * < 0.05. ASCL1, Achaete-Scute Family BHLH Transcription Factor 1; CHGA, Chromogranin A; ePNEC, epithelial-derived pulmonary neuroendocrine cells; iPNEC, iPSC-derived pulmonary neuroendocrine cells; iPSC, induced pluripotent stem cells; SYP, Synaptophysin.

Article Snippet: Cells were grown using the bronchial epithelial cell growth kit from ATCC (PCS300- 040).

Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Derivative Assay, Control, RNA Sequencing, MANN-WHITNEY

Journal: Allergy

Article Title: Human Pulmonary Neuroendocrine Cells Respond to House Dust Mite Extract With PAR-1 Dependent Release of CGRP.

doi: 10.1111/all.16416

Figure Lengend Snippet: FIGURE 4 | CGRP release after HDM stimulation is PAR1-dependent (A) Single-cell transcriptomics data of iPNECs (GSE146990, Hor et al.) and ePNEC for PAR1 (F2R), PAR2 (F2RL1), PAR3 (F2RL2), and PAR4 (F2RL3) genes. (B) SYP and PAR1 immunohistochemical co-staining in naive iPNEC (male, 32). 20× and 100× magnification and scale bars at 50 and 10 μm, respectively. (C, D) CALCB mRNA (C) and CGRP protein (D) expression in different ePNEC (● male, 52; ▲ male, 56; ○ female, 55) conditions after 2 h: 50 μL PBS, HDM (1200 μg/mL), PAR1 inhibitor Vorapaxar (80 μM) and PAR1 agonist TFLLR-NH2 (2 μM). (E) CALCB mRNA RT-PCR expression in different ePNEC conditions after 2 h: 50 μL PBS, HDM (1200 μg/mL), HDM co-incubation with protease inhibitors Chymostatin (broad spectrum, 10 μg/mL), PMSF (serine specific, 0.25 mM) and E-64 (cysteine specific, 10 μM). Inhibitor/agonist and or HDM added at the same time. Each dot represents one well and data shown for mean ± SEM. One-way ANOVA for (C, D, E); * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. Symbol on top of multiple brackets represents all brackets. Agonist, PAR1 agonist vorapaxar; ePNEC, epithelial-derived pulmonary neuroendocrine cells; Inh., PAR1 inhibitor; iPNEC, iPSC-derived pulmonary neuroendocrine cells; HDM, House dust mite; PAR1, Protease activated receptor 1; SYP, Synaptophysin.

Article Snippet: Cells were grown using the bronchial epithelial cell growth kit from ATCC (PCS300- 040).

Techniques: Single-cell Transcriptomics, Immunohistochemical staining, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Incubation, Derivative Assay

Journal: Molecular cancer research : MCR

Article Title: Defects in Emerin-nucleoskeleton binding disrupt nuclear structure and promote breast cancer cell motility and metastasis

doi: 10.1158/1541-7786.MCR-20-0413

Figure Lengend Snippet: (A) Western Blot analysis of emerin in primary mammary epithelial cells, MCF10A cells, MDA-231 cells and MDA-157 cells. (B) Western Blot quantification of emerin expression for each cell line. Emerin expression was normalized to tubulin and primary mammary epithelial cells. Error bars represent standard error. (n=3) ****p-value < 0.0001, unpaired t-test. (C) Nuclear area for MCF10A (n=116, blue), MDA-231 (n=237, green) and MDA-157 (n=342, red) cells. Error bars represent standard error. ****p-value < 0.0001, unpaired t-test (D) Representative DAPI (blue) images of MCF10A, MDA-231, and MDA-157 cells. Scale bars: 100 μm. (E) Nuclear circularity for MCF10A (n=75, blue), MDA-231 (n=301, green), and MDA-157 (n=237, red) cells. Error bars represent standard error. ****p-value < 0.0001, unpaired t-test

Article Snippet: Primary mammary epithelial cells were grown in mammary epithelial cell basal medium (ATCC, cat#: PCS-600–030) with all the components from the mammary epithelial cell growth kit (ATCC, cat#: PCS-600–040).

Techniques: Western Blot, Expressing