Journal: bioRxiv

Article Title: PDFGRα + Stromal Cells Promote Salivary Gland Proacinar Differentiation Through FGF2-dependent BMP7 Signaling

doi: 10.1101/2021.11.19.469144

Figure Lengend Snippet: A) Transcriptome heatmap showing enrichment of specific acinar (green) and ductal (salmon) genes with FGF2 (lavender) and control (cyan) organoid culture conditions, respectively. Log two-fold scale for gene expression is shown with red (higher) and blue (lower). B) GSEA plots depicting microarray differential gene expression lists compared with ductal and acinar gene lists. Plots show FGF2 cultured organoids enriched for acinar genes, while control organoids are enriched in ductal genes. Enrichment score (ES) and nominal p-value are provided. C) E16 SMG organoids grown with or without stroma and with or without exogenous FGF2 were immunostained to detect Calponin 1 (CNN1, green) with the nuclear stain DAPI (blue). Only organoids grown with stromal cells contain CNN1 + epithelium. Scale: 200 µm. D) Quantification of organoid area positive for CNN1 normalized to DAPI. Organoids made with stroma have significantly more myoepithelium; mean±s.d n=4 technical replicates, Single-factor ANOVA with post-huc Tukey test. E) Immunostained E16 SMG epithelial and stromal cell-containing organoids cultured with and without FGF2. An EPCAM + (green) and CNN1 + (red) myoepithelial layer is present within in the epithelial compartment surrounded by HSPG2 + (white) basement membrane (arrowheads) both in the presence and absence of FGF2. Nuclei stained with DAPI (blue). Scale: 50 µm.

Article Snippet: The stroma was negatively selected against epithelium using a EPCAM antibody coated microbeads (1:10; Miltenyi Biotec #130-061-101) and a Miltenyi MS column.

Techniques: Control, Gene Expression, Microarray, Cell Culture, Staining, Membrane

Journal: bioRxiv

Article Title: PDFGRα + Stromal Cells Promote Salivary Gland Proacinar Differentiation Through FGF2-dependent BMP7 Signaling

doi: 10.1101/2021.11.19.469144

Figure Lengend Snippet: A) E16 organoids created using epithelium cultured with: FGF2 and no stroma (FGF2 Only), stroma without FGF2 (Stroma Only), stroma and FGF2 (Stroma & FGF2) or with PDGFRα + MACS-isolated stroma and FGF2 (PDGFRα Stroma & FGF2). ICC for EPCAM (white) shows the epithelium, PDGFRα + (red) shows stromal subset, and DAPI (blue) shows nuclei. All condition form epithelial organoids, however stromal PDGFRα only increases when FGF2 is present. B) PDGFRα + positive area was quantified relative to DAPI. PDGFRα + MACS-selected stroma shows a >2-fold increase in PDGFRα similar to the total stroma when FGF2 is present; mean±s.d n=4, 5, 5, 4 experimental replicates, Single factor ANOVA with post-hoc Tukey Test. C) E16 organoids created using epithelium cultured with: FGF2 and no stroma (FGF2 Only), stroma without FGF2 (Stroma Only), stroma and FGF2 (Stroma & FGF2) or with PDGFRα + MACS-isolated stroma and FGF2 (PDGFRα + Stroma & FGF2). ICC for AQP5 (green) shows the proacinar epithelium and DAPI (blue) shows nuclei. AQP5 increases equally when either total stroma or PDGFRα + stroma are combined with FGF2. D) AQP5 positive area was quantified relative to DAPI. AQP5 increased when either total stroma or PDGFRα + stroma are in the presence of FGF2 containing media; mean±s.d n=11, 9, 14, and 5 experimental replicates, Single factor ANOVA with post-huc Tukey Test. E) Organoids created using E16 epithelium cultured in FGF2 media and combined with: no stroma, E16 total stroma, or adult PDGFRα + MACS-selected stroma. ICC showing proacinar epithelium using AQP5 (green) and DAPI (blue). The adult PDGFRα + stroma also support proacinar epithelium similarly to the total E16 stroma. F) APQ5 positive area quantified relative to DAPI. The organoids cultured with either E16 stroma or adult PDGFRα + stroma had a 3-fold higher AQP5 + relative area than organoids cultured without stroma; mean±s.d n=4 technical replicates, Single-factor ANOVA with post-hoc Tukey Test.

Article Snippet: The stroma was negatively selected against epithelium using a EPCAM antibody coated microbeads (1:10; Miltenyi Biotec #130-061-101) and a Miltenyi MS column.

Techniques: Cell Culture, Isolation

Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Validation of the top genes by TaqMan RT-PCR in the cell lines.

Article Snippet: EPCAM , Hs00158980_m1 , 201839_s_at , epithelial cell adhesion molecule , , 0.009.

Techniques: Biomarker Discovery, Binding Assay

Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Representative examples of the immunhistochemical validation for CD9, EpCAM and LGALS8. Left column: normal kidney, right column: selected tumor tissue.

Article Snippet: EPCAM , Hs00158980_m1 , 201839_s_at , epithelial cell adhesion molecule , , 0.009.

Techniques: Biomarker Discovery

Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Kaplan-Meier survival plots of sunitinib-treated metastatic RCC samples divided into two cohorts based on the median of EpCAM positive cells (p = 0.01).

Article Snippet: EPCAM , Hs00158980_m1 , 201839_s_at , epithelial cell adhesion molecule , , 0.009.

Techniques: