Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Chlamydia trachomatis induces the transcriptional activity of host YAP in a Hippo-independent fashion

doi: 10.3389/fcimb.2023.1098420

Figure Lengend Snippet: Chlamydia infection induces expression of a subset of YAP target genes. (A) Volcano plot of gene expression in bulk RNA-sequencing of End1/E6E7 immortalized epithelial cells (End1s) during infection with Chlamydia trachomatis ( Ct ) serovar L2 compared to mock infection, at 24 hours post-infection (hpi). n = 3, with a minimum of 3x10 7 unstranded single reads per replicate with a mean length of 150 bp. All fold changes are relative to the mock-infected control; red dots: false discovery rate p-value (FDRP) ≤ 0.05, labels: top 20 genes whose expression differed most significantly (lowest FDRP) from the mock infection. (B) Table of selected transcription factors identified as potential targets of infection-associated modulation via cross-referencing of differentially expressed genes identified in (A) with the ChIP Enrichment Analysis (ChEA) database of transcription factor target genes. See also Supplementary Data S2. (C) Volcano plot of gene expression in bulk RNA-sequencing of primary human endocervical epithelial cells (HCECs) during infection with Ct serovar L2 compared to mock infection, at 24 hours post-infection (hpi). n = 3, with a minimum of 3x10 7 unstranded single reads per replicate with a mean length of 150 bp. All fold changes are relative to the mock-infected control; red dots: false discovery rate p-value (FDRP) ≤ 0.05, labels: top 20 genes whose expression differed most significantly (lowest FDRP) from the mock infection. (D) Venn diagram of differentially expressed genes identified in (A, C) cross-referenced with the ChEA database of YAP target genes. (E) Scatter plot of gene expression of YAP-responsive (ChEA), differentially expressed genes in either Ct serovar L2-infected End1s (x-axis) or HCECs (y-axis). All fold changes are relative to each cell type’s respective mock-infected control; blue line: linear regression model of correlation; grey shading: 95% confidence interval. R 2 and p-values calculated using Pearson’s correlation. (F) Heatmap of YAP target gene expression in Ct serovar L2-infected End1s (left columns) and HCECs (right columns). All fold changes are relative to each cell type’s respective mock-infected control; only genes differentially expressed (FDRP ≤ 0.05) in both cell types are shown. (G) Dot plot of GO biological process term enrichment in the set of YAP target genes differentially expressed in Ct serovar L2 infection of End1s or HCECs (top 25 most significantly enriched terms shown). Dot size: number of term-associated genes found in set, dot color: adjusted p-value.

Article Snippet: Primary human cervical epithelial cells (HCECs, ATCC PCS-0480-011, Lot 80306190) were cultured at 37° C with 5% atmospheric CO 2 in Cervical Epithelial Cell Basal Medium (CECBM, ATCC PCS-480-032) supplemented with all contents of a Cervical Epithelial Growth Kit (ATCC PCS-080-042).

Techniques: Infection, Expressing, Gene Expression, RNA Sequencing, Control, Targeted Gene Expression

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Chlamydia trachomatis induces the transcriptional activity of host YAP in a Hippo-independent fashion

doi: 10.3389/fcimb.2023.1098420

Figure Lengend Snippet: Chlamydia infection promotes YAP nuclear translocation. (A) Expression of CTGF and CYR61 at 24 hpi in mock- and Ct L2-infected End1 cells, as measured by RT-qPCR. n = 3 biological replicates; fold changes are relative to mean expression of the mock-infected and untreated control. Whiskers: minimum to maximum; n.s.: not significant (p-values > 0.05), asterisks: p-values ≤ 0.05, using pairwise Student’s t-tests and Bonferroni’s correction for multiple comparisons. (B) Expression of CTGF at 24 hpi in mock- and Ct L2-infected End1 cells transfected with non-targeting (NT) or YAP1-targeting siRNA (10 nM for 24 h prior to infection), as measured by RT-qPCR. n = 3 biological replicates; fold changes are relative to mean expression of the mock-infected and untreated control. Whiskers: minimum to maximum; asterisks: p-values ≤ 0.05, using pairwise Student’s t-tests and Bonferroni’s correction for multiple comparisons. (C) Expression of CTGF, INHBA, and BMP2 at 24 hpi in mock- and Ct L2-infected End1 cells treated with the YAP-TEAD inhibitor verteporfin (Vpf, 5 μM for 16 h starting at 8 hpi), as measured by RT-qPCR. n = 5 biological replicates; fold changes are relative to mean expression of the mock-infected and untreated control. Whiskers: minimum to maximum; asterisks: p-values ≤ 0.05, using pairwise Student’s t-tests and Bonferroni’s correction for multiple comparisons. (D) Representative micrographs of YAP (green) translocation into the nuclei (blue) of confluent mock- and Ct L2-infected End1 cells at 24 hpi. Asterisks: chlamydial inclusions, scale bar: 20 μm. (E) Quantification of YAP nuclear translocation in (D) as a ratio of nuclear to cytosolic YAP fluorescence. n = 5 biological replicates, 50 cells measured per sample. Black bars: group means; asterisks: p-values ≤ 0.05, using pairwise Wilcoxon rank sum tests and Bonferroni’s correction for multiple comparisons. (F) Representative micrographs of YAP nuclear translocation of confluent mock- and Ct L2-infected primary human cervical epithelial cells at 24 hpi. Asterisks, chlamydial inclusions, scale bar: 20 μm. (G) Quantification of YAP nuclear translocation in (F) as a ratio of nuclear to cytosolic YAP fluorescence. n = 5 biological replicates, 50 cells measured per sample. Black bars: group means; asterisks: p-values ≤ 0.05, using pairwise Wilcoxon rank sum tests and Bonferroni’s correction for multiple comparisons. (H) Quantification of YAP nuclear translocation at 2, 4, 8, 12, 18, and 24 hpi in confluent mock- and Ct L2-infected End1 cells as a ratio of nuclear to cytosolic YAP fluorescence. n = 5 biological replicates, 50 cells measured per sample. Blue dots: mock-infected cells, red dots: Ct L2-infected cells, black bars: group means, asterisks: p-value ≤ 0.05, using pairwise Wilcoxon rank-sum tests and Bonferroni’s correction for multiple comparisons. (I) Representative micrographs of YAP nuclear translocation at 18 hpi in confluent mock- and Ct L2-infected End1 cells treated with chloramphenicol (Cm, 50 μg/mL for 1 h at 17 hpi) or DMSO. Asterisks: chlamydial inclusions; scale bar: 20 μm. (J) Quantification of YAP nuclear translocation in (I) . n = 3 biological replicates, 50 cells measured per sample. Black bars: group means; asterisks: p-values ≤ 0.05, using pairwise Wilcoxon rank-sum tests and Bonferroni’s correction for multiple comparisons.

Article Snippet: Primary human cervical epithelial cells (HCECs, ATCC PCS-0480-011, Lot 80306190) were cultured at 37° C with 5% atmospheric CO 2 in Cervical Epithelial Cell Basal Medium (CECBM, ATCC PCS-480-032) supplemented with all contents of a Cervical Epithelial Growth Kit (ATCC PCS-080-042).

Techniques: Infection, Translocation Assay, Expressing, Quantitative RT-PCR, Control, Transfection, Fluorescence

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Airway branching generations in human and rat investigated in this study included BG0-6 in humans and BG0-5 in rats. b Workflow for imaging luminal epithelial cell type composition and ciliary beat and clearance function in airway samples. Schematic: Created in BioRender. Nawroth, J. (2025) https://BioRender.com/a01i578 ). c Example IF staining of cilia (ATUB, magenta) and secretory cells (SCGB1A1, green; MUC5AC, gray) in human and rat airway epithelium in BG0 (trachea) and BG5/6. Scale bar: 20 µm. d Quantification of luminal cell proportions labeled with ATUB (ciliated cells) or with MUC5AC and/or SCGB1A1 (secretory cells) as a function of airway branching generation in human and rat airway epithelium. Inset: Percentage of human secretory cell population positive for only MUC5AC (gray), only for SCGB1A1 (green), or for both (white) as a function of branching generation. Solid line: mean, shaded region: SEM. Numbers of human donors (2–3 FOVs each): BG0, n = 3; BG1, n = 3; BG2, n = 5; BG3, n = 3; BG4, n = 7; BG5, n = 2; BG6, n = 2; Numbers of rat donors (2–3 FOVs each): BG0, n = 5; BG1, n = 3; BG2, n = 2; BG4, n = 1; BG5, n = 1. For full donor information see Supplementary Tables and . Source data for ( d ) are provided as a file.

Article Snippet: SAGM: Small Airway Epithelial Growth Medium (Promocell, C-21170) containing 50 nM EC23; and 5. mAir: a 1:1 mix of Dulbecco’s modified eagle medium and Airway epithelial cell growth medium (AECGM, PromoCell, C-21160) with AECGM supplements and 50 nM EC23 (previously described in ref. ).

Techniques: Imaging, Staining, Labeling

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Representative measurement of ciliary beat frequency (CBF) and associated particle clearance trajectories and speed in a human airway epithelial sample (BG2). b Same measurements in rat airway epithelial sample (BG1). Scalebar in ( a , b ): 100 µm. c Quantification of average CBF, particle clearance speed, and clearance per beat (CPB) in human airways BG0-6 and rat airways BG0-1. Number of human donors: n = 4. Number of rat donors: n = 6. d left: Clearance directionality as a function of distance in human (red) and rat (blue) airways. Thick lines are average curves. Right: Mean directionality over a flow distance of 80 µm. Number of human donors n = 4; number of rat donors: n = 4. Boxplots: Each solid dot is the mean value of one donor (1–3 BGs, 2–4 FOVs each); red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum; significance was assessed with two-sided unpaired t-test. Source data for ( c , d ) are provided as a file.

Article Snippet: SAGM: Small Airway Epithelial Growth Medium (Promocell, C-21170) containing 50 nM EC23; and 5. mAir: a 1:1 mix of Dulbecco’s modified eagle medium and Airway epithelial cell growth medium (AECGM, PromoCell, C-21160) with AECGM supplements and 50 nM EC23 (previously described in ref. ).

Techniques:

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Representative IF-images of primary human airway epithelial cultures from 1 donor grown in different differentiation media for 28 days at ALI and stained for cilia (ATUB, magenta) and secretory cell markers (SCGB1A1, green; MUC5AC, white). Scalebar, 40 µm. b Average luminal cell type composition based on IF-staining (in vitro: N = 4–7 donors, 2 inserts each with 3–8 FOVs each; ex vivo: 9 donors, 1–3 BGs, 2–3 FOVs each). c Mapping of IF-staining data of in vitro and ex vivo samples onto three dimensions. y- axis: percentage of MUC5AC+ cells; x-axis: cilia coverage, i.e., percentage of ciliated (ATUB + ) cells; circle diameter: ratio of SCGB1A1+ to MUC5AC+ cell percentages. d Average percentage of MUC5B expressing cells and e relative percentage of MUC5B expressing cells that also express either SCGB1A1 or MUC5AC from IF-stainings (in vitro: n = 2 (SAGM, BD) to n = 4 (PC, PC-S, mAir) donors, 1–2 inserts each with 1–6 FOVs each; ex vivo: n = 3 donors, BG0, 2–4 FOVs each). Boxplot: Each dot represents mean of one donor; red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum. Source data for ( b – e ) are provided as a file.

Article Snippet: SAGM: Small Airway Epithelial Growth Medium (Promocell, C-21170) containing 50 nM EC23; and 5. mAir: a 1:1 mix of Dulbecco’s modified eagle medium and Airway epithelial cell growth medium (AECGM, PromoCell, C-21160) with AECGM supplements and 50 nM EC23 (previously described in ref. ).

Techniques: Staining, In Vitro, Ex Vivo, Expressing

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Quantitative analysis of particle CPB and directionality in primary human airway epithelial cultures grown in different differentiation media for 28 days at ALI compared to human and rat benchmark data. Number of donors (2–3 inserts each with 6 FOVs each) for BD, mAir, SAGM, PC, PC-S, respectively: n = 5, 4, 4, 5, 4. b Quantitative analysis of ciliary beat metrics in airway cells cultured and visualized as in ( a ). Number of donors (2–3 inserts each with 6 FOVs each) for BD, mAir, SAGM, PC, PC-S, respectively: Cilia Coverage, n = 7, 4, 4, 7, 4; Ciliary Beat OP, n = 4, 4, 2 (1 not measurable), 4, 4; Ciliary Beat Amplitude: n = 4, 4, 3, 4, 4; Cilia Length, n = 4, 4, 4, 4, 4; Ciliation Gap, n = 4, 4, 3, 4, 4; Crystalline OP, n = 4, 4, 3, 4, 4. Boxplots in ( a , b ): Each solid dot is the mean value of one donor (1–3 BGs, 2–4 FOVs each); red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum. Dotted lines indicate average human and rat benchmark values. c Top: Predicted CPB and clearance directionality in in vitro cultures compared to predicted human airway performance (red). Shaded regions indicate uncertainty based on spread of input metrics. Measured mean values per donor are overlaid (dots). Bottom: Predicted change in CPB and directionality of in vitro cultures in different media if an individual cilia input parameter is set to match ex vivo values. Source data for ( a – c ) are provided as a file.

Article Snippet: SAGM: Small Airway Epithelial Growth Medium (Promocell, C-21170) containing 50 nM EC23; and 5. mAir: a 1:1 mix of Dulbecco’s modified eagle medium and Airway epithelial cell growth medium (AECGM, PromoCell, C-21160) with AECGM supplements and 50 nM EC23 (previously described in ref. ).

Techniques: Cell Culture, In Vitro, Ex Vivo

Journal: bioRxiv

Article Title: Development of high-affinity, single-domain protein binders for neutralizing household allergens

doi: 10.1101/2025.08.03.668213

Figure Lengend Snippet: (a) Thermostability profile of AVA-1-1-C3 as a representative protein, as determined by differential scanning fluorimetry. (b-g) Cytotoxicity of primary human cells following 48-hour exposure to AVA-1-1-C3 or an equivalent concentration of bovine serum albumin (BSA), evaluated using the CellTiter-Glo® (CTG) luminescent cell viability assay. Cell types included human epithelial keratinocytes (HEK, b), human dermal fibroblasts (HDF, c), human follicle dermal papilla cells (HFDPC, d), human renal cortical epithelial cells (HRCEPC, e), human skeletal muscle cells (HSKMC, f), and human dermal microvascular endothelial cells (HDMEC, g). No significant differences in cell viability were observed. Data in (b-g) represents mean ± s.d. and are representative of two experimental replicates.

Article Snippet: Cells were cultured in their corresponding complete media including HDF growth medium (Cell Applications, Cat# 116-500), human EpiVita serum-free growth medium (Cell Applications, Cat# 141-500a), microvascular endothelial cell growth medium (PromoCells, Cat# C-22120), HSkMC growth medium (Cell Applications, Cat# 151-500), and renal epithelial cell growth media (PromoCell, Cat# C-26130).

Techniques: Concentration Assay, Cell Viability Assay

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Airway branching generations in human and rat investigated in this study included BG0-6 in humans and BG0-5 in rats. b Workflow for imaging luminal epithelial cell type composition and ciliary beat and clearance function in airway samples. Schematic: Created in BioRender. Nawroth, J. (2025) https://BioRender.com/a01i578 ). c Example IF staining of cilia (ATUB, magenta) and secretory cells (SCGB1A1, green; MUC5AC, gray) in human and rat airway epithelium in BG0 (trachea) and BG5/6. Scale bar: 20 µm. d Quantification of luminal cell proportions labeled with ATUB (ciliated cells) or with MUC5AC and/or SCGB1A1 (secretory cells) as a function of airway branching generation in human and rat airway epithelium. Inset: Percentage of human secretory cell population positive for only MUC5AC (gray), only for SCGB1A1 (green), or for both (white) as a function of branching generation. Solid line: mean, shaded region: SEM. Numbers of human donors (2–3 FOVs each): BG0, n = 3; BG1, n = 3; BG2, n = 5; BG3, n = 3; BG4, n = 7; BG5, n = 2; BG6, n = 2; Numbers of rat donors (2–3 FOVs each): BG0, n = 5; BG1, n = 3; BG2, n = 2; BG4, n = 1; BG5, n = 1. For full donor information see Supplementary Tables and . Source data for ( d ) are provided as a file.

Article Snippet: Cells were then seeded at a density of 30 K cells cm -2 on PureCol (Advanced Biomatrix #5005) coated tissue culture dishes in airway epithelial growth media (Promocell #C-21160) and passaged at 80% confluence.

Techniques: Imaging, Staining, Labeling

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Representative measurement of ciliary beat frequency (CBF) and associated particle clearance trajectories and speed in a human airway epithelial sample (BG2). b Same measurements in rat airway epithelial sample (BG1). Scalebar in ( a , b ): 100 µm. c Quantification of average CBF, particle clearance speed, and clearance per beat (CPB) in human airways BG0-6 and rat airways BG0-1. Number of human donors: n = 4. Number of rat donors: n = 6. d left: Clearance directionality as a function of distance in human (red) and rat (blue) airways. Thick lines are average curves. Right: Mean directionality over a flow distance of 80 µm. Number of human donors n = 4; number of rat donors: n = 4. Boxplots: Each solid dot is the mean value of one donor (1–3 BGs, 2–4 FOVs each); red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum; significance was assessed with two-sided unpaired t-test. Source data for ( c , d ) are provided as a file.

Article Snippet: Cells were then seeded at a density of 30 K cells cm -2 on PureCol (Advanced Biomatrix #5005) coated tissue culture dishes in airway epithelial growth media (Promocell #C-21160) and passaged at 80% confluence.

Techniques:

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Representative IF-images of primary human airway epithelial cultures from 1 donor grown in different differentiation media for 28 days at ALI and stained for cilia (ATUB, magenta) and secretory cell markers (SCGB1A1, green; MUC5AC, white). Scalebar, 40 µm. b Average luminal cell type composition based on IF-staining (in vitro: N = 4–7 donors, 2 inserts each with 3–8 FOVs each; ex vivo: 9 donors, 1–3 BGs, 2–3 FOVs each). c Mapping of IF-staining data of in vitro and ex vivo samples onto three dimensions. y- axis: percentage of MUC5AC+ cells; x-axis: cilia coverage, i.e., percentage of ciliated (ATUB + ) cells; circle diameter: ratio of SCGB1A1+ to MUC5AC+ cell percentages. d Average percentage of MUC5B expressing cells and e relative percentage of MUC5B expressing cells that also express either SCGB1A1 or MUC5AC from IF-stainings (in vitro: n = 2 (SAGM, BD) to n = 4 (PC, PC-S, mAir) donors, 1–2 inserts each with 1–6 FOVs each; ex vivo: n = 3 donors, BG0, 2–4 FOVs each). Boxplot: Each dot represents mean of one donor; red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum. Source data for ( b – e ) are provided as a file.

Article Snippet: Cells were then seeded at a density of 30 K cells cm -2 on PureCol (Advanced Biomatrix #5005) coated tissue culture dishes in airway epithelial growth media (Promocell #C-21160) and passaged at 80% confluence.

Techniques: Staining, In Vitro, Ex Vivo, Expressing

Journal: Nature Communications

Article Title: Structure and function relationships of mucociliary clearance in human and rat airways

doi: 10.1038/s41467-025-57667-z

Figure Lengend Snippet: a Quantitative analysis of particle CPB and directionality in primary human airway epithelial cultures grown in different differentiation media for 28 days at ALI compared to human and rat benchmark data. Number of donors (2–3 inserts each with 6 FOVs each) for BD, mAir, SAGM, PC, PC-S, respectively: n = 5, 4, 4, 5, 4. b Quantitative analysis of ciliary beat metrics in airway cells cultured and visualized as in ( a ). Number of donors (2–3 inserts each with 6 FOVs each) for BD, mAir, SAGM, PC, PC-S, respectively: Cilia Coverage, n = 7, 4, 4, 7, 4; Ciliary Beat OP, n = 4, 4, 2 (1 not measurable), 4, 4; Ciliary Beat Amplitude: n = 4, 4, 3, 4, 4; Cilia Length, n = 4, 4, 4, 4, 4; Ciliation Gap, n = 4, 4, 3, 4, 4; Crystalline OP, n = 4, 4, 3, 4, 4. Boxplots in ( a , b ): Each solid dot is the mean value of one donor (1–3 BGs, 2–4 FOVs each); red line indicates median, bottom and top edges of the box indicate 25th and 75th percentiles, whiskers indicate minimum and maximum. Dotted lines indicate average human and rat benchmark values. c Top: Predicted CPB and clearance directionality in in vitro cultures compared to predicted human airway performance (red). Shaded regions indicate uncertainty based on spread of input metrics. Measured mean values per donor are overlaid (dots). Bottom: Predicted change in CPB and directionality of in vitro cultures in different media if an individual cilia input parameter is set to match ex vivo values. Source data for ( a – c ) are provided as a file.

Article Snippet: Cells were then seeded at a density of 30 K cells cm -2 on PureCol (Advanced Biomatrix #5005) coated tissue culture dishes in airway epithelial growth media (Promocell #C-21160) and passaged at 80% confluence.

Techniques: Cell Culture, In Vitro, Ex Vivo