Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Characterization of PRLR isoform antibodies. A. Western blot analysis indicating specificity of polyclonal antibodies. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Cell lysates were prepared and proteins separated by PAGE. Each isoform specific lysate was probed with each isoform specific antibody. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. Molecular weights are marked as indicated: LF = 93 kDa, SF1a = 57 kDa, SF1b = 38 kDa. Cell lysates from CHO transfected cells were immunoprecipitated with their respective antibodies, then immunoblotted with the same antibodies. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. IP indicates immunoprecipitation. B. Western blot analysis using commercially available PRLR antibody. ECD = antibody recognizing extracellular domain C. Fluorescent immunocytochemical analysis. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Specific staining was observed using isoform specific polyclonal antibodies. The negative control (not shown) lacks primary antibody and appears black. Data shown are representative of triplicate experiments. Magnification 20×.

Article Snippet: PRLR isoform expression was examined on 12 lobular carcinoma and 10 ductal carcinoma specimens obtained from CHTN; other investigators may have received samples from these same tissues.

Techniques: Western Blot, Transfection, Immunoprecipitation, Staining, Negative Control

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunoanalysis on human xenografts. A. Fluorescent immunohistochemistry was performed on MCF7, MDA-MB-231, or T47D cells which were injected into the cleared fat pads of nude mice; once tumors reached 1 cm 2 (measured in two dimensions), they were excised, formalin fixed, sectioned, and stained with the indicated PRLR isoform specific antibodies. B. Immunohistochemical analysis using DAB as chromogen of MCF7, MDA-MB-231, or T47D xenograft tumors. neg = no primary antibody as negative control; DAPI stain to visualize the presence of cells. Photographs are representative from 5 different tumors for each cell line. Magnification 40×.

Article Snippet: PRLR isoform expression was examined on 12 lobular carcinoma and 10 ductal carcinoma specimens obtained from CHTN; other investigators may have received samples from these same tissues.

Techniques: Immunohistochemistry, Injection, Staining, Immunohistochemical staining, Negative Control

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Message for PRLR isoforms in breast carcinomas. RNA was extracted from 7 ductal (left) and 7 lobular (right) carcinomas and qPCR was performed. Data shown are the mean +/- SE of delta delta Ct values relative to SF1a expression.

Article Snippet: PRLR isoform expression was examined on 12 lobular carcinoma and 10 ductal carcinoma specimens obtained from CHTN; other investigators may have received samples from these same tissues.

Techniques: Expressing

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunohistochemical analysis from ductal breast tissue arrays. Representative photographs taken of ductal carcinoma specimens to demonstrate absent, low, medium, or high amounts of fluorescent red intensity staining. 144 individual ductal specimens were stained with PRLR isoform specific antibodies and analyzed. Scale bar indicates range of red scoring.

Article Snippet: PRLR isoform expression was examined on 12 lobular carcinoma and 10 ductal carcinoma specimens obtained from CHTN; other investigators may have received samples from these same tissues.

Techniques: Immunohistochemical staining, Staining

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunohistochemical analysis from lobular breast tissue arrays. Representative photographs of lobular carcinoma specimens to demonstrate absent, low, medium, or high amounts of fluorescent red intensity staining. 104 individual lobular specimens were stained with PRLR isoform specific antibodies and analyzed. Scale bar indicates range of red scoring.

Article Snippet: PRLR isoform expression was examined on 12 lobular carcinoma and 10 ductal carcinoma specimens obtained from CHTN; other investigators may have received samples from these same tissues.

Techniques: Immunohistochemical staining, Staining

Journal: bioRxiv

Article Title: Inhibition of the gut ceramidase Asah2 decelerates the vertebrate ageing rate

doi: 10.64898/2026.04.30.721799

Figure Lengend Snippet: a , Schematic representation of ceramide hydrolysis by membrane-tethered Asah2. Cer, ceramide; Sph, sphingosine. Image was created with BioRender.com. b , Loss of Asah2 activity is associated with a longer lifespan. Protein domain architectures of Asah2 are shown. Blue box, catalytic domain; orange box, immunoglobulin (Ig)-like domain; white box, other regions; grey box, N. furzeri MZM-specific region; red vertical lines in human ASAH2, essential amino acid residues required for enzymatic activity. c, e , asah2 is specifically expressed in the intestine and increases with age. Expression levels of asah2 mRNA in various tissues ( c ) and in the intestine ( e ) of 2- and 4-month-old wild-type (WT) and asah2 knockout (KO) N. furzeri are shown. In c and e , mRNA levels were analysed by reverse transcription quantitative PCR (RT–qPCR) and RNA sequencing (RNA-seq), respectively. CPM, counts per million; FDR, false discovery rate. FDR values were derived from quasi-likelihood methods in edgeR. d , Asah2 protein localises to the apical /luminal side of intestinal epithelium in short-lived GRZ fish but is absent in long-lived MZM fish and asah2 KO fish. Representative images of intestinal sections immunostained with anti-Asah2 antibody (green) and 4′,6-diamidino-2-phenylindole (DAPI; blue) are shown. Scale bar, 200 μm. f , Pooled censoring-aware parental survival analysis showing that higher participant plasma ASAH2 z-score was associated with higher pooled parental hazard, consistent with lower parental longevity. The displayed p-value tests the overall ASAH2-parental hazard association in the stratified cubic B-spline Cox model. Primary statistical inference was based on the cluster-robust linear Cox model. g , CRISPR/Cas9-mediated asah2 knockout. Top, schematic representation of the asah2 locus (black boxes, exons; ATG, start codon) and two sgRNA target sites (orange boxes). Bottom left, DNA sequences of WT and asah2 KO. The red horizontal line denotes the 4 bp deletion. Bottom right, the mutation was predicted to result in loss of the C-terminal domain of the Asah2 protein. Blue, catalytic domain; orange, Ig-like domain; white, other regions; grey, asah2 KO N. furzeri- specific region. h , Expression of Asah2 in the intestine of short-lived N. furzeri analysed by Western blotting. i, j , asah2 KO extends lifespan ( i ) and improves locomotor activity ( j ). In i , survival curves of male WT (blue) and asah2 KO (orange) fish are shown on the left; the corresponding statistical results, mean lifespan and maximum lifespan are shown on the right. Black dots indicate censored individuals. Graphs in j show exploratory behaviour in male WT (blue) and asah2 KO (orange) fish. The y-axis indicates the average velocity. The log-rank test, fixed-time survival test, and unpaired two-tailed t -test were used for i , and a Brown–Forsythe and Welch ANOVA followed by Dunnett’s T3 multiple-comparisons test were used for j . Bars and error bars represent the mean ± SD.

Article Snippet: Human tissue expression data were obtained from the Human Protein Atlas , .

Techniques: Membrane, Activity Assay, Expressing, Knock-Out, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, RNA Sequencing, Derivative Assay, Clinical Proteomics, CRISPR, Mutagenesis, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Inhibition of the gut ceramidase Asah2 decelerates the vertebrate ageing rate

doi: 10.64898/2026.04.30.721799

Figure Lengend Snippet: a , Asah2 expression increases with age in the human terminal ileum of the small intestine. Data were obtained from voyAGEr . CPM, counts per million. b , Sex-stratified pooled censoring-aware parental survival analysis of participant plasma ASAH2 z-score and pooled parental hazard in male and female participants. Displayed p-values denote the overall ASAH2-parental hazard association from stratified cubic B-spline Cox models. c, d , asah2 KO does not affect body size or body mass index ( c ) or reproductive capacity ( d ). Graphs in c show body weight, body length, and body mass index in WT (blue) and asah2 KO (orange) male fish. Animals measured at each time point were different individuals. In d , the graph shows the number of fertilized eggs in WT (blue) and asah2 KO (orange) fish. Bars and error bars ( c, d ) represent the mean ± SD. An unpaired two-tailed t -test was used in c , and a Brown–Forsythe and Welch ANOVA followed by Dunnett’s T3 multiple-comparisons test were used in d .

Article Snippet: Human tissue expression data were obtained from the Human Protein Atlas , .

Techniques: Expressing, Clinical Proteomics, Two Tailed Test

Journal: Nature Communications

Article Title: Combinatory differentiation of human induced pluripotent stem cells generates functional thymic epithelium driving dendritic cell and CD4/CD8 T cell development

doi: 10.1038/s41467-026-68675-y

Figure Lengend Snippet: a Reaggregation of iPSc-derived TEPs with primary human ETPs to form hTOs (human Thymic Organoids) cultured in a 3D fibrin hydrogel at an air-liquid interface in medium supplemented with RANKL, IL7, SCF, and FLT3L (Created in BioRender. Guillonneau, C. (2025) https://BioRender.com/apk8n61 ). b Bright-field images of hTOs (human Thymic Organoids) at 24 h and D4 showing a compact spheroid core with a peripheral cellular crown (identical magnification) ( n = 12 from independent differentiations and ETP donors). c scRNA-seq of EPCAM + CD45 − cells from D7 organoids with UMAP embedding and cell-cycle scoring (10x Genomics; Seurat workflow). d Module score for a TEP gene set: PDPN , PAX9 , IL7 , EBF1 , FN1 . e Integration of the EPCAM + CD45 − scRNA-seq dataset (Clusters 1, 2, and 3) with the Bautista human TEC atlas; with reference cells in gray. f Cell type deconvolution of TEPs and CD205 + TEPs against the EPCAM + CD45 − scRNA-seq dataset. g Distribution of the Tau tissue-specific variability index within clusters 3 and 1. Tau was computed from GTEx tissue expression data; TRA enrichment was assessed on the top-200 cluster-specific genes. Source data are provided as a file. h Assessment of the TRA repertoire in Cluster 3, showing enrichment for brain antigens. Source data are provided as a file.

Article Snippet: The Tau was calculated for a list of genes using the gene tissue expression data (GTEx) from the Human Protein Atlas database ( https://www.proteinatlas.org/about/download ).

Techniques: Derivative Assay, Cell Culture, Expressing