Journal: International Journal of Molecular Sciences

Article Title: The Impact of Alternate-Day Fasting on the Salivary Gland Ductal Compartments and the Differentiation Potential of Keratin 5 + Salivary Gland Progenitor Cells in an Induced Mouse Model of Sjögren’s-like Hyposalivation

doi: 10.3390/ijms27094080

Figure Lengend Snippet: Immunization with salivary gland proteins induces hyposalivation, accompanied by elevated P16 expression in the submandibular glands (SMGs) of KRT5 CreERT2 ; R26 tdTomato mice. Eight-week-old female KRT5 CreERT2 ; R26 tdTomato mice were immunized with total salivary gland proteins following tamoxifen treatment. ( A ) Experimental timeline of tamoxifen induction, immunizations and analyses. ( B ) Measurement of salivary flow rate (n = 6–8). ( C ) Representative images of immunohistochemical staining of the SMG sections for P16 (scale bar = 100 μm, 400× magnification). The bar graph shows the average percentage of positively stained areas (n = 4–5). Error bars indicate the standard error of the mean (SEM).

Article Snippet: Female KRT5 CreERT2 mice (Cat# 029155), R26 tdTomato reporter mice (Cat# 007914) and C57BL/6 mice (Cat# 000664) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained under specific pathogen-free conditions at the ADA Forsyth Institute.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Alternate-Day Fasting on the Salivary Gland Ductal Compartments and the Differentiation Potential of Keratin 5 + Salivary Gland Progenitor Cells in an Induced Mouse Model of Sjögren’s-like Hyposalivation

doi: 10.3390/ijms27094080

Figure Lengend Snippet: ADF increases salivary flow rate and reduces cellular senescence in the SMGs of KRT5 CreERT2 ; R26 tdtomato mice with induced hyposalivation. Eight-week-old female KRT5 CreERT2 ; R26 tdTomato mice were immunized with SMG proteins following tamoxifen treatment and then subjected to alternate-day fasting (ADF) or fed with standard chow ad libitum (AL), as shown in ( A ) experimental timeline. ( B ) Measurement of salivary flow rate (n = 6–8). ( C ) Representative images of immunohistochemical staining of the SMG sections for P16 (scale bar = 100 μm, 400× magnification). The bar graph shows the average percentage of positively stained areas (n = 5–6). Error bars indicate the standard error of the mean (SEM).

Article Snippet: Female KRT5 CreERT2 mice (Cat# 029155), R26 tdTomato reporter mice (Cat# 007914) and C57BL/6 mice (Cat# 000664) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained under specific pathogen-free conditions at the ADA Forsyth Institute.

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Alternate-Day Fasting on the Salivary Gland Ductal Compartments and the Differentiation Potential of Keratin 5 + Salivary Gland Progenitor Cells in an Induced Mouse Model of Sjögren’s-like Hyposalivation

doi: 10.3390/ijms27094080

Figure Lengend Snippet: ADF reduces the expression of anti-apoptotic proteins BCL-2, BCL-XL and MCL-1 in the SMGs of KRT5 CreERT2 ; R26 tdtomato mice with induced hyposalivation. Eight-week-old female KRT5 CreERT2 ; R26 tdTomato mice were immunized with SMG proteins following tamoxifen treatment and then subjected to ADF or fed with standard chow AL , as outlined in A. Representative images of immunohistochemical staining of the SMG sections for BCL-2, BCL-XL or MCL-1 (scale bar = 100 µm, 400× magnification). The bar graphs display the average percentage of positively stained areas (n = 5–6). Error bars indicate the standard error of the mean (SEM).

Article Snippet: Female KRT5 CreERT2 mice (Cat# 029155), R26 tdTomato reporter mice (Cat# 007914) and C57BL/6 mice (Cat# 000664) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained under specific pathogen-free conditions at the ADA Forsyth Institute.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Alternate-Day Fasting on the Salivary Gland Ductal Compartments and the Differentiation Potential of Keratin 5 + Salivary Gland Progenitor Cells in an Induced Mouse Model of Sjögren’s-like Hyposalivation

doi: 10.3390/ijms27094080

Figure Lengend Snippet: ADF reduces the expression levels of NLRP3 and their downstream products IL-1β and IL-18 in the SMGs of KRT5 CreERT2 ; R26 tdTomato mice with induced hyposalivation. Eight-week-old female KRT5 CreERT2 ; R26 tdTomato mice were immunized with SMG proteins following tamoxifen treatment and then subjected to ADF or fed with standard chow AL , as outlined in A. Representative images of immunohistochemical staining of the SMG protein sections for NLRP3, IL-1β and IL-18 are shown (scale bar = 100 µm, 400× magnification). The bar graphs display the average percentage of positively stained areas (n = 5–6). Error bars indicate the standard error of the mean (SEM).

Article Snippet: Female KRT5 CreERT2 mice (Cat# 029155), R26 tdTomato reporter mice (Cat# 007914) and C57BL/6 mice (Cat# 000664) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained under specific pathogen-free conditions at the ADA Forsyth Institute.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Alternate-Day Fasting on the Salivary Gland Ductal Compartments and the Differentiation Potential of Keratin 5 + Salivary Gland Progenitor Cells in an Induced Mouse Model of Sjögren’s-like Hyposalivation

doi: 10.3390/ijms27094080

Figure Lengend Snippet: ADF moderately promotes acinar cell differentiation from KRT5 + cells in the SMGs of KRT5 CreERT2 ; R26 tdTomato mice with induced hyposalivation. ( A ) Flow cytometric analysis of KRT5 and tdTomato expression in SMG cells from KRT5 CreERT2 ; R26 tdTomato mice 2 weeks after corn oil or tamoxifen treatment. ( B ) Eight-week-old female KRT5 CreERT2 ; R26 tdTomato mice were immunized with SMG proteins following tamoxifen treatment and then subjected to ADF or fed with standard chow AL , as outlined in A. Representative images of immunofluorescence staining of the SMG sections for tdTomato (red) and AQP5 (acinar cell marker, green) are shown. DAPI: blue. Scale bar = 100 µm, 400× magnification (n = 9). The arrow indicates cell aggregates co-expressing tdTomato and AQP5.

Article Snippet: Female KRT5 CreERT2 mice (Cat# 029155), R26 tdTomato reporter mice (Cat# 007914) and C57BL/6 mice (Cat# 000664) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained under specific pathogen-free conditions at the ADA Forsyth Institute.

Techniques: Cell Differentiation, Expressing, Immunofluorescence, Staining, Marker

Journal: bioRxiv

Article Title: Comparative analysis of varicella-zoster virus and herpes simplex virus 1 interaction with epidermal terminal differentiation in primary human keratinocytes models of differentiation

doi: 10.64898/2026.04.08.717198

Figure Lengend Snippet: A. Schematic of plan of infection with HSV-1 at MOI 0.005 of undifferentiated (Undiff.), early differentiated (Early diff.) and late differentiated (Late diff.) NHEKs. “Undiff.”, “Early diff.” and “Late diff.” labelling refers to the state of NHEKs differentiation at the time of infection (day 0). The “Undiff.” condition includes the NHEKs undergoing spontaneous asynchronous differentiation after infection (Differentiating_async.) B. Analysis by qPCR of cell-associated HSV-1 genome copy number normalised to number of cells, upon infection with HSV-1 of Undiff., Early diff. and Late diff. NHEKs, and evaluated at 24h, 48h and 72h p.i. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control) and are reported as mean of n=3 independent experiments ± SEM. C. Analysis by qPCR of cell-associated HSV-1 genome copy number normalised to number of cells, upon infection with HSV-1 of Late diff. NHEKs, and reported at 72h and 120h p.i. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control) and are reported as the mean of three technical replicates. D. ICP0 (pink) immunofluorescence staining of Undiff., Early diff. and Late diff. NHEKs infected with HSV-1 and analysed at 4h, 24h and 72h p.i. The reported images include GFP (green) signal deriving from the GFP protein tagged to the late HSV-1 tegument protein UL46 and DAPI (grey) staining of nuclei. E. Analysis by qRT-PCR analysis of RL2 gene (encoding the ICP0 protein) and UL48 gene (encoding the VP16 protein) expression in HSV-1 infection of Undiff., Early diff. and Late diff. NHEKs, and reported at 24h, 48h and 72h p.i. The data are reported as mean of 2 -dCT of n=3 independent experiments ± SEM, where the normalisation was performed against GAPDH . F. Western Blotting analysis of ICP0 and VP16 expression at 24h, 48h and 72h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with HSV-1. GAPDH was used as loading control. Data are representative of n=4 independent experiments (Undiff. condition), n=3 independent experiments (Early diff. condition) and n=2 independent experiments (Late diff. condition). G. Schematic of the model of HSV-1 infection implemented in undifferentiated NHEKs, which either remained undifferentiated throughout the infection and until the last time point (72h) (Undifferentiated) or started undergoing spontaneous differentiation during the last time points of infection (Differentiating_async). H. Analysis by qRT-PCR of KRT5 and KRT10 mRNAs in uninfected NHEKs at 72h confirming the undifferentiated and differentiating status of the keratinocytes. The data are reported as fold change (FC) (2 -ddCT ) to the undifferentiated condition and are the mean of three technical replicates, where the normalisation was performed against GAPDH . I. Keratinocytes where infected when undifferentiated and then either maintained their undifferentiated status throughout the infection (Undifferentiated) or were allowed to spontaneously differentiate by 72h p.i. (Differentiating). Cell-associated HSV-1 genome copy number was analysed at 72h p.i. in both conditions. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control), are reported as the mean of three technical replicates and are representative of n=3 independent experiments. J. Anlysis by qRT-PCR of RL2 and UL48 mRNAs in HSV-1 infected Undifferentiated and Differentiating NHEKs at 72h p.i. The data are reported as mean of 2 -dCT of three technical replicates, where the normalisation was performed against GAPDH . Data are representative of n=3 independent experiments. Statistical significance was evaluated in B. and E. by 2way ANOVA with Tukey’s multiple comparisons test and indicated as *P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001. In D. , dotted lines mark nuclei at 4h p.i. Insets contain zoomed-in areas of the ICP0 signal inside the nuclei at 4h p.i. Scale bar, 10 μm. Undiff., undifferentiated; Early diff., early differentiated; Late diff., late differentiated; p.i., post infection; NHEKs, normal human epidermal keratinocytes.

Article Snippet: The primers and probes used for host genes were purchased from Thermofisher Scientific, cat n 4448489 (VIC dye): GAPDH (ASSAY ID Hs02786624_g1) and cat n 4331182 (FAM dye): KRT10 (assay ID Hs00166289_m1), KRT1 (assay ID Hs00196158_m1), KRT5 (assay ID Hs00361185_m1), LOR (assay ID Hs01894962_s1), FLG (assay ID Hs00856927_g1), LCE3D (assay ID Hs00754375_s1), DSC1 (assay ID Hs00245189_m1).

Techniques: Infection, Generated, Control, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing, Western Blot

Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: a Representative images of DCLK1 and BCAM staining in tracheal sections from the indicated Sox9-tdT reporter strains at day 14. Sox9-tdT + BCAM + cells (white arrow). Scale bars: 50 µm. b Quantification of Sox9-tdT + BCAM + cells in the SAE from the indicated strains at day 14. Group sizes: n = 5, 6, 12, 5 mice, from left to right. Two-tailed unpaired Mann-Whitney U test. c , d Combined in situ hybridization (ISH) for Oxgr1, Krt5 and Krt8 , and immunofluorescence for SOX9 and α-SMA in tracheal sections from WT mice with or without ALT treatment ( Krt5 for BCs, Krt8 for differentiated cells, SOX9 for SMG progenitor cells, and α-SMA for SMG myoepithelial cells). Oxgr1 + SOX9 + cells (green arrows). Scale bars: 20 μm. SMG Submucosal gland. ( e ) Quantification of Oxgr1 + cells in the specified EpC subsets. Group sizes: n = 7, 16, 8, 13, 6, 10, 7, 8 mice, from left to right. Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction. f Quantification of Oxgr1 + cells among the Krt5 + or Krt8 + cells. Group sizes: n = 12, 11, 12, 7 mice, from left to right. Two-tailed unpaired Mann-Whitney U test. g Experimental schema. h , i Representative images ( h ) and quantification ( i ) of BCAM and SOX9 staining in tracheal sections from the indicated strains at day 14. Scale bars: 20 μm. Group sizes: n = 5, 8, 7, 5 mice, from left to right. Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction. The dashed line marks the border between SAE and SM. Box plots ( b , e , f , i ) show median (center line), 25th–75th percentiles (box), and min–max values (whiskers). Each data point represents the average of 3–4 images from one mouse. See “Methods”. Panel g was created in BioRender. Lee, M. ( https://BioRender.com/dg3i7rm ). Source data are provided as a Source Data file.

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques: Staining, Two Tailed Test, MANN-WHITNEY, In Situ Hybridization, Immunofluorescence

Journal: Regenerative Therapy

Article Title: Airway basal stem cell derived extracellular vesicles promote lung repair in chronic obstructive pulmonary disease

doi: 10.1016/j.reth.2026.101068

Figure Lengend Snippet: Lineage tracing of BC–mediated repair in the tracheal epithelium after LPS&PPE injury. (a) Schematic of the Krt5 Cre ERT2 -Gt (ROSA)26 Sor tm4(ACTB-tdTomato-EGFP) reporter system used for lineage tracing of airway basal stem cells. (b) Experimental timeline illustrating tamoxifen induction, LPS&PPE-induced emphysema injury, and subsequent analysis at designated time points. (c) Representative immunofluorescence images of tracheal sections at different time points before and after injury. GFP (green) marks lineage-labeled basal cells and their progeny. Co-staining with Krt5 (red) and p63 (red) identifies basal cells, with CC10 (red) and Foxj1 (red) marking Club and ciliated cells, respectively. Scale bar, 20 μm. (d) Quantification of lineage-labeled cells categorized as basal, Club, or ciliated cells at the indicated time points before and after injury. Data are presented as mean ± SEM (n = 1–2 technical replicates from 3 mice). (e) Representative immunofluorescence images of lung tissue sections showing GFP + (green) lineage-labeled cells and Krt5 (red) basal cells distributed along proximal–distal airways at different time points before and after injury. Scale bar, 100 μm.

Article Snippet: Krt5 Cre ERT2 -Gt (ROSA)26Sor tm4(ACTB-tdTomato-EGFP) mice (The Jackson Laboratory, USA) were used to perform lineage tracing.

Techniques: Immunofluorescence, Labeling, Staining

Journal: Regenerative Therapy

Article Title: Airway basal stem cell derived extracellular vesicles promote lung repair in chronic obstructive pulmonary disease

doi: 10.1016/j.reth.2026.101068

Figure Lengend Snippet: Intratracheal transplantation of BCs promotes alveolar repair in emphysematous mice. (a) Representative bright-field image of clonogenic BCs cultured on feeder layers. Scale bar, 100 μm. (b) Immunofluorescence staining of clonogenic BCs using Krt5 (green) and p63 (red) as basal cell markers. Scale bars, 100 μm. (c) Schematic diagram illustrating the experimental design for intratracheal transplantation of BCs into mice pre-treated with BLM&PPE or LPS&PPE. (d) Representative direct fluorescence image of recipient mouse lung 7 days after intratracheal delivery of 10 6 GFP-labeled BCs. (e) Immunofluorescence staining of lung sections showing engraftment and distribution of transplanted GFP + BCs (green), some of which retained Krt5 expression (red). Scale bar, 200 μm. (f) Representative H&E staining of lung sections from healthy control mice and emphysematous mice treated with PBS or BCs. Emphysema was induced by intratracheal administration of LPS and PPE. Scale bar 50 μm. (g) Quantification of average alveolar count, mean alveolar area, and percentage of lung injury across the three groups. Data are presented as mean ± SEM (n = 3 mice). Statistical analysis was performed using a one-way ANOVA with Tukey's multiple comparisons test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Krt5 Cre ERT2 -Gt (ROSA)26Sor tm4(ACTB-tdTomato-EGFP) mice (The Jackson Laboratory, USA) were used to perform lineage tracing.

Techniques: Transplantation Assay, Cell Culture, Immunofluorescence, Staining, Fluorescence, Labeling, Expressing, Control