a agr2  (Cell Signaling Technology Inc)

Journal: EMBO Molecular Medicine

Article Title: Benchmarking porcine pancreatic ductal organoids for drug screening applications

doi: 10.1038/s44321-025-00330-3

Figure Lengend Snippet: ( A – C ) Single-plane confocal images of PPDO derived from one Em pig pancreas immunostained against Pan-Cytokeratin (PAN-CK)-BMPR1A-CDH1 ( A ), SOX9-Phalloidin-NKX6-1 ( B ), CFTR-GP2-PDX1 ( C ) and counterstained with DAPI. Insets show the individual channels of the merge image. PPDO were stained at passage 2. Scale bar: 50 µm. ( D –F ) Single-plane confocal images of PPDO derived from an LPN pig pancreas immunostained against KRT5-BMPR1A-CDH1 ( D ), SOX9-GCG/NKX6-1-Phalloidin ( E ), CFTR-GP2-PDX1 ( F ) and counterstained with DAPI. Insets show the individual channels of the merge image. PPDO were stained at passage 4. Scale bar: 50 µm. ( G – I ) Single-plane confocal images of PPDO derived from an Ad pig pancreas immunostained against CFTR-GP2-PDX1 ( G ), SOX9-Phalloidin-CDH1 ( H ), AMY3A-KRT7-NEUROG3 ( I ), and counterstained with DAPI. PPDO were stained at passage 3. Insets show the individual channels of the merge image. Scale bar: 50 µm. ( J –L ) Single-plane confocal images of HPDO immunostained against KRT7-AGR2-CDH1 ( J ), CFTR-BMPR1A-NKX6.1 ( K ), SOX9-Phalloidin-PDX1 ( L ), and counterstained with DAPI. Insets show the individual channels of the merge image. HPDO were stained at passage 3. Scale bar: 50 µm. ( M , N ) Single-plane confocal images of porcine pancreas cryosections (Ad) immunostained against NKX6.1-SOX9-CDH1 ( M ) and phalloidin-555-PDX1 ( N ), and counterstained with DAPI. Scale bar: 50 µm. ( O ) Quantification of the organoid area of late passage (>5) PPDO following 4 h of live imaging and treatment with forskolin. n = 3 independent experiments with 3 PPDO lines (2 EPN and 1 Ad). Data are shown as mean ± SD. One-way ANOVA followed by Dunnett’s multiple comparisons test was used to assess significance. Not-significant (ns) P = 0.9010.

Article Snippet: Primary antibodies against KRT7 (mouse, 1:200, Agilent Technologies M701829-2) and AGR2 (rabbit, 1:200, Cell Signaling 13062S) were used.

Techniques: Derivative Assay, Staining, Imaging

Journal: EMBO Molecular Medicine

Article Title: Benchmarking porcine pancreatic ductal organoids for drug screening applications

doi: 10.1038/s44321-025-00330-3

Figure Lengend Snippet: ( A , B ) UMAP representation of the integrated PPDO and primary porcine pig ductal and β-cells scRNA-Seq datasets (2 LPN pancreata) with the corresponding annotated cell types/states ( A ) as well as the corresponding individual samples used for integration ( B ). sysVI integration is shown. ( C – E ) UMAP representation of the pig pancreas dataset showing the corresponding cell types ( E ) and the expression level of the LGR5 ( F ) and AGR2 ( G ) genes. Arrowheads point to the LGR5 and AGR2 positive ductal cell population. ( F ) Single-plane confocal images of EPN porcine pancreas immunostained against KRT7 (gray) and AGR2 (magenta) and counterstained with DAPI. Insets show the individual channels of the merge image. Scale bar: 50 µm. ( G ) Single-plane confocal images of PPDO immunostained against KRT7 (gray) and AGR2 (magenta) and counterstained with DAPI. Insets show the individual channels of the merge image. Scale bar: 50 µm. ( H , I ) Brightfield images of PPDO cultured in medium supplemented with ( H ) or without ( I ) WNT3a/R-spondin-1/Noggin factors (WRN) 7 days following duct isolation and culture. Scale bar 500 µm. PS = Passage. ( J , K ) Brightfield images of PPDO cultured in medium supplemented with ( J ) or without ( K ) WNT3a/R-spondin-1/Noggin factors (WRN) after three passages at 7 days of culture post passage. Scale bar 500 µm. Experiments in ( H – K ) have been independently reproduced using PPDO from three different animals (1 EPN and 2 Ad). ( L , M ) Brightfield images of HPDO cultured in medium supplemented with ( L ) or without ( M ) WNT3a/R-spondin-1/Noggin factors (WRN) 7 days following duct isolation and culture. Scale bar 500 µm. ( N , O ) Brightfield images of HPDO cultured in medium supplemented with ( N ) or without ( O ) WNT3a/R-spondin-1/Noggin factors (WRN) after three passages at 7 days of culture post passage. Scale bar 500 µm. Experiments in ( N , O ) have been independently reproduced using 4 different HPDO preparations. .

Article Snippet: Primary antibodies against KRT7 (mouse, 1:200, Agilent Technologies M701829-2) and AGR2 (rabbit, 1:200, Cell Signaling 13062S) were used.

Techniques: Expressing, Cell Culture, Isolation