Journal: bioRxiv

Article Title: Endometrial Hyperplasia Risk Is Increased by High-Fat Diet Via Estrogen-Driven Stromal Fibroblast Reprogramming Toward a Pro-Fibrotic State

doi: 10.64898/2026.03.20.713224

Figure Lengend Snippet: (a,b) Representative H&E stain of endometrial tissue from Pten heterozygous mutants for each diet. Main image= 10x magnification; inset=20x magnification; scalebar=300 μm. Caliper indicates the height of an abnormal endometrial gland. (c,d) Representative KRT8 IHC stain of endometrial tissue from Pten heterozygous mutants for each diet (CD n=5; HFD n=4). Main image= 10x magnification; inset=20x magnification; scalebar=300 μm. (e,f) Representative PTEN IHC stain of endometrial tissue from Pten heterozygous mutants for each diet (CD n=9; HFD n=11). Main image= 20x magnification; inset=40x magnification; scalebar=200 μm. Arrow indicates a gland with PTEN loss, while an arrowhead indicates a PTEN intact gland. (g,h) Representative Masson’s Trichrome stain of endometrial stromal ECM from Pten heterozygous mutants for each diet. Main image= 20x magnification; inset=40x magnification; scalebar=200 μm. Arrow indicates endometrial stroma. (i-m) Graphs quantifying EH disease severity in either CD- (n=10) or HFD- (n=11) treated non-estrous staged Pten heterozygous mutant mice from H&E-stained endometrial slides. Measured disease severity parameters were total number of glands per representative histological section, percent normal glands (%), percent abnormal glands (%), average abnormal gland epithelial height (μm), and average abnormal gland area (μm). Datapoints represent a value for each mouse (bar=mean ± s.d.). Two-tailed unpaired t -test statistic with Welch’s correction as needed. (n-q) Graphs quantifying ECM raw integrated density or fold change in integrated density in Pten heterozygous mutant endometrial stroma using Image J with the Color Deconvolution plugin. Samples were from either estrus-staged or non-estrous staged mice in both CD- (estrus n=5; non-estrus n=10) or HFD- (estrus n=4; non-estrus n=11) treated animals. Measurement (datapoint) represents each mouse’s mean of 4 samplings (bar=mean ± s.d.). Two-tailed unpaired t -test statistic. (r) Percent (%) of CD45 + cells from all live single cells in the uterus of CD (n=5) and HFD (n=8) Pten heterozygous mice. Measurement (datapoint) represents each mouse (bar=mean ± s.d.). Two-tailed unpaired t -test statistic. (s) Percent (%) of F4/80 + cells from all CD45 + live single cells in the uterus. Measurement (datapoint) represents each mouse (bar=mean ± s.d.). Two-tailed unpaired t -test statistic with Welch’s correction. (t,u) Percent (%) of dead cells or CD45 + immune cells from all single cells in blood of CD (n=5) and HFD (n=7) Pten heterozygous mice. Measurement (datapoint) represents each mouse (bar=mean ± s.d.). Two-tailed unpaired t -test statistic. (v) Percent (%) of F4/80 + cells from CD45 + single cells in the blood. Blood F4/80 + cells are likely monocytes, as macrophages are not commonly seen in circulation. Measurement (datapoint) represents each mouse (bar=mean ± s.d.). Two-tailed unpaired t -test statistic. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001

Article Snippet: Samples were labelled 1:50 CD45 (Miltenyi, cat#130-123-784, RRID:AB_2802059), 1:50 Propidium Iodide (PI) (BD Biosciences, cat#556463, RRID:AB_2869075), 1:50 F4/80 (BioLegend, cat#123120, RRID:AB_893479), 1:20 CD80 (BioLegend, cat#104713, RRID:AB_313134), 1:40 CD206 (BioLegend, cat#141707, RRID:AB_10896057), as well as isotype controls at a matching concentration including APC Rat IgG2b,τχ (BioLegend, cat#403806, RRID:AB_ 3096358), Alexa Fluor® 488 Rat IgG2a, κ (BioLegend, cat#400525, RRID:AB_2864283), APC Armenian Hamster IgG (BioLegend, cat#400911, RRID:AB_2905474), and APC Rat IgG2a, κ (BioLegend, cat#400511, RRID:AB_2814702).

Techniques: Staining, Mutagenesis, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: A BLM intratracheal model scheme (n = 3 per group). B H&E and Masson validation of inflammation/fibrosis (n = 3 per group). C Lung scRNA-seq UMAP. D scATAC-seq UMAP subclusters. E Venn showing Piezo1 and Piezo2 as co-regulated genes across silica and BLM bulk, scRNA-seq and scATAC-seq data. F scATAC-seq coverage peaks of Piezo1 and Piezo2 . G CD31 and CD45-sorted EC bulk-RNA-seq heatmap and bar chart (n = 3 per group). Gene expression levels were Z-score normalized. P values were calculated using the two-tailed unpaired t-test. H PIEZO1 (green) and Ve-cad (red) co-staining and quantification in fibrotic mouse lungs (n = 4). UMAP and bar charts (mean ± SEM) of PIEZO1 + ( I ) and PIEZO2 + ( J ) EC counts in normal controls (n = 5) versus IPF (n = 4) patients. P values were calculated using the two-tailed unpaired t-test. K PIEZO1 (green) and CD31 (red) co-staining and quantification in human IPF vs normal lungs (n = 10). L Meta-analysis of five GEO datasets showing elevated PIEZO1 in IPF ECs. Standardized mean difference (SMD) were calculated as Hedges’ g with corresponding 95% confidence intervals. Data were shown as mean ± SEM, H and K: two-tailed unpaired t-tests, Source data are provided as a Source Data file.

Article Snippet: Harvested mouse lungs were washed with pre-cooled DPBS, cut into pieces, digested with digestion DMEM containing 1 mg/mL type I collagenase (Solarbio, #C8140), 1 mg/mL type IV collagenase (Biosharp, C#8160), 1 mg/mL Dispase II (Solarbio, #D6430), and 0.2 mg/mL DNAase (Solarbio, #BS165) in 37 °C for 40 min. After erythrocytes were lysed, cells were resuspended with MACS Buffer (Miltenyi Biotec, #130-091-376) and incubated on ice for 15 minutes with anti-mouse CD31 MicroBeads (for endothelial cell separation, Miltenyi Biotec, #130-097-418), anti-mouse CD45 MicroBeads (for immune cell separation, Miltenyi Biotec, #130-052-301), anti-mouse CD326 MicroBeads (for epithelial cell separation, Miltenyi Biotec, #130-105-958), and anti-mouse CD140a MicroBeads (for fibroblast separation, Miltenyi Biotec, #130-101-502).

Techniques: Biomarker Discovery, RNA Sequencing, Gene Expression, Two Tailed Test, Staining