Journal: Nature Communications

Article Title: Pulmonary fibroblast activation during Aspergillus fumigatus infection enhances lung defense via immunomodulation and tissue remodeling

doi: 10.1038/s41467-026-71027-5

Figure Lengend Snippet: A Schematic representation of the TAM-inducible Cre recombinase system. Cre recombinase, flanked by a tamoxifen-inducible modified estrogen receptor ( MerCreMer ), was inserted into the Postn locus, enabling temporal control over Cre activity in cells that activate the Postn promoter. In the presence of TAM, Cre translocates to the nucleus and catalyzes recombination between loxP sites flanking a transcriptional stop upstream of a tdTomato reporter gene inserted into the rosa26 locus. Cre activity results in constitutive expression of tdTomato in Cre-expressing cells and their progeny, thereby fluorescently marking Postn Lin fibroblasts. B Schematic representation of the experimental approach used to study fibroblast activation in Postn Lin immunocompetent mice after OP inoculation with 3 × 10 7 A. fumigatus conidia. TAM was administered via gavage one day before the fungal challenge and maintained through a TAM-infused chewing diet for the duration of the experiment. C Representative fluorescent photomicrographs showing the induction of tdTomato-positive cells following exposure to the specified treatments. Arrows indicate sporadic Postn Lin cells in samples not treated with A. fumigatus . D , E Time course illustrating the appearance of tdTomato + cells on days 1, 3, 5, and 7 post-inoculation with A. fumigatus . Bars represent the means ± SD from 3 biological replicates, each shown as an individual dot (* p < 0.05; one-way ANOVA on ranks with Tukey’s post hoc test). F Flow cytometric analysis of fold-change in tdTomato expression following inoculation with 3 × 10 7 conidia relative to mock infection with saline. Values represent the means ± SD from 4 and 5 biological replicates in the control and A. fumigatus -treated groups, respectively. Dots indicate individual biological replicates (* p < 0.05; unpaired two-tailed t -test). Source data are provided as a Source Data file.

Article Snippet: Rosa26-tdTomato (B6.Cg- Gt(rosa)26Sor tm14(CAG-tdTomato)Hze /J; JAX Strain #:007914), Rosa26-DTA (B6.129S6(Cg)- Gt(ROSA))26Sor tm1(DTAJpmb /J; JAX Strain #:032087), Pdgfrα-CreERT2 (B6.129S- Pdgfra tm1.1(cre/ERT2)Blh /J; JAX Strain #:032770), and Pdgfrα-eGFP (B6.129S4- Pdgfra tm11(EGFP)Sor /J; JAX Strain #:007669) mice were purchased from The Jackson Laboratory.

Techniques: Modification, Control, Activity Assay, Expressing, Activation Assay, Infection, Saline, Two Tailed Test

Journal: Nature Communications

Article Title: Pulmonary fibroblast activation during Aspergillus fumigatus infection enhances lung defense via immunomodulation and tissue remodeling

doi: 10.1038/s41467-026-71027-5

Figure Lengend Snippet: A Schematic of the TAM-inducible DTA ablation model. The Cre recombinase, inserted into the Postn locus, is controlled by a TAM-inducible estrogen receptor ( MerCreMer ). Upon Postn promoter activation, TAM-induced recombination of the DTA gene results in DTA toxin expression from the rosa26 locus, preventing fibroblast activation and expansion. B Schematic representation of the experimental approach: Mice were inoculated OP with 1 × 10 6 conidia, followed by immunosuppression with triamcinolone acetonide 5 h post-infection. TAM was administered via gavage one day before fungal inoculation and maintained through a TAM-infused chewing diet for the duration of the experiment. C Percent survival of immunosuppressed mice infected with A. fumigatus ( n = 24 for control group and n = 20 for DTA group; p = 3.8 × 10 −5 , log rank test). D Gross appearance of lungs from infected DTA and control mice on day 4 post-infection. E Representative histopathologic images showing focal alveolar hemorrhage and angioinvasion in the A. fumigatus -infected DTA mice by H&E and GMS staining, respectively. F Distribution of alveolar hemorrhage severity in control (white bars) and DTA mice (red bars). Alveolar hemorrhage was scored semi quantitatively as described in Fig. . Data are shown as box-and-whisker plots with individual biological replicates overlaid ( n = 10 for control group and n = 8 for DTA group). Boxes represent the interquartile range (IQR), horizontal lines indicate the median, and whiskers extend to 1.5 x IQR (two-tailed Mann–Whitney U test). G Quantification of angioinvasion in intrapulmonary arteries of control (white bars) and DTA (red bars) mice. The percentage of vessels ≥100 µm in diameter that contained A. fumigatus hyphae in each animal were quantified in GMS-stained lung sections. Values represent the mean ± SD from 10 and 8 biological replicates in the control and DTA groups, respectively, with individual replicates indicated by dots (unpaired two-tailed t -test). H Collagen deposition quantified from Picrosirius Red-stained sections of lung tissue. Values represent the mean ± SD from 10 and 8 biological replicates in the control (white bars) and DTA (red bars) groups, respectively, with individual replicates indicated by dots (ns, no significant; unpaired two-tailed t -test) ( I ) Schematic representation of the experimental approach: similar to the schematic shown in ( B ), but lung tissue was harvested at day 4 post-challenge for further analysis, coinciding with the highest mortality rate observed in ( C ). J Flow cytometric quantification of viable Pdgfrα + cells. Right panels show the cell gating strategy used to distinguish viable and dead cells within the Pdgfrα + population. Values represent the mean ± SD from 5 and 8 biological replicates in the control and DTA groups, respectively ( p = 3.5 × 10 −5 ; unpaired two-tailed t -test). K Fungal burden quantification by RT-qPCR from lung lobes of the indicated mouse models. Fungal burden is expressed as Genome Equivalents (GE) per mg of lung tissue. Values represent the mean ± SD from 5 and 8 biological replicates in the control (white bars) and DTA (red bars) groups, respectively, with individual replicates indicated by dots ( p < 0.05; unpaired two-tailed t -test). L Quantification of immune infiltration by flow cytometry. Total leukocytes and hematopoietic subsets were discriminated using the antibodies indicated on the y -axis. Values represent the mean ± SD from 5 and 8 biological replicates in the control and DTA groups, respectively, with individual replicates indicated by dots (ns, no significant, unpaired two-tailed t-test). Source data are provided as a Source Data file.

Article Snippet: Rosa26-tdTomato (B6.Cg- Gt(rosa)26Sor tm14(CAG-tdTomato)Hze /J; JAX Strain #:007914), Rosa26-DTA (B6.129S6(Cg)- Gt(ROSA))26Sor tm1(DTAJpmb /J; JAX Strain #:032087), Pdgfrα-CreERT2 (B6.129S- Pdgfra tm1.1(cre/ERT2)Blh /J; JAX Strain #:032770), and Pdgfrα-eGFP (B6.129S4- Pdgfra tm11(EGFP)Sor /J; JAX Strain #:007669) mice were purchased from The Jackson Laboratory.

Techniques: Activation Assay, Expressing, Infection, Control, Staining, Whisker Assay, Two Tailed Test, MANN-WHITNEY, Quantitative RT-PCR, Flow Cytometry

Journal: Investigative Ophthalmology & Visual Science

Article Title: Oligodendrocyte Precursor Cells Shape Retinogeniculate Refinement Via a CHD8-Dependent Phagocytic Pathway

doi: 10.1167/iovs.67.3.64

Figure Lengend Snippet: Depletion of OPCs impairs synaptic elimination in dLGN. ( A ) Top : Diagram of Olig2 -TK mouse generation. Bottom : Schematic of OLIG2 + mitotic cells ablation. ( B ) Left : OLIG2 immunostaining in the dLGN of P14 wild-type (Ctrl) and Olig2 -TK mice. Scale bar = 100 µm. Right : Quantification of OLIG2 + cells. ( C ) Left : Immunolabeling for Synapsin + pre-synapses around MAP2 + neurons in dLGN of P14 Ctrl and Olig2 -TK mice. Scale bar = 50 µm. Right : Quantification of Synapsin + signal around a single neuron. ( D ) Left : Representative images of vGLUT2 and HOMER1 immunostaining in the dLGN of P14 Ctrl and Olig2 -TK mice. Scale bar = 25 µm. Right : Quantification of vGLUT2 + /HOMER1 + synapse puncta. ( E ) Left : Immunostaining for vGAT in dLGN of P14 Ctrl and Olig2 -TK mice. Scale bar = 50 µm. Right : Quantification of percent vGAT + area. ( F ) Top : Diagram of tamoxifen (TAM) administration induced DTA expression. Bottom : Schematic of TAM treatment from P3 to P7 for OPC depletion. ( G ) Left : PDGFRα immunolabeling in dLGN of P14 PDGFRα-Cre ERT (Ctrl) and PDGFRα-Cre ERT : DTA ( Pα -iDTA) mice. Scale bar = 100 µm. Right : Quantification of PDGFRα + cells. ( H ) Left : Immunolabeling for Synapsin + presynapses around MAP2 + neurons in the dLGN of P14 Ctrl and Pα -iDTA mice. Scale bar = 50 µm. Right : Quantification of Synapsin + signal around a single neuron. ( I ) Left : Representative images of vGLUT2 and HOMER1 immunostaining in the dLGN of P14 Ctrl and Pα -iDTA mice. Scale bar = 25 µm. Right : Quantification of vGLUT2 + /HOMER1 + synapse puncta. ( J ) Left : vGAT immunolabeling in the dLGN of P14 Ctrl and Pα -iDTA mice. Scale bar = 50 µm. Right : Quantification of percent vGAT + area ( n = 5 animals per genotype). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 and n.s., not significant (unpaired, Two-tailed Student's t -test).

Article Snippet: PDGFRα-Cre ERT BAC mice were crossed with DTA mice (JAX, 009669) to generate the Pα -iDTA.

Techniques: Immunostaining, Immunolabeling, Expressing, Two Tailed Test