Journal:

Article Title: Bone-Marrow-Derived Cells Contribute to Glomerular Endothelial Repair in Experimental Glomerulonephritis

doi:

Figure Lengend Snippet: Immunofluorescent double-staining of glomerular sections from WRBM→BN rats at 28 days after induction of anti-Thy-1.1 nephritis. Confocal laser-scanning microscopy (×630, pinhole 2.5). A: Staining for BM donor (WR) MHC-I (U9F4, green). B: Staining for endothelial cells (RECA-1, red). C: U9F4/RECA-1 double-staining identifies bone-marrow-derived endothelial cells, randomly integrated in the glomerular endothelium (U9F4/ED-1 double-positive, yellow). D: Staining for BM donor (WR) MHC-I (U9F4, green). E: Staining for mesangial cells (OX-7, red). F: U9F4/OX-7 double-staining showing donor bone-marrow-derived mesangial cells (U9F4/OX-7 double-positive, yellow) integrated in the glomerulus. G: Staining for BM donor (WR) MHC-I (U9F4, red). H: Staining for monocytes/macrophages (ED-1, green). I: U9F4/ED-1 double-staining shows a BM donor-derived (U9F4/ED-1 double positive, yellow) monocyte/macrophage. ▵▵, U9F4/OX-7, U9F4/RECA-1, U9F4/ED-1 double-positive cells (yellow), ie, donor-derived mesangial and endothelial cells and a donor-derived monocyte/macrophage. ∧∧, single-U9F4 staining.

Article Snippet: The following primary antibodies were used: fluorescein isothiocyanate (FITC)-labeled anti-BN-MHC-I (CL027F; Cedarlane, Hornby, Ontario, Canada), anti-WR-MHC-I (U9F4 19 ; kindly provided by J. Rozing, Groningen, the Netherlands), and secondary antibody goat anti-mouse-phycoerythrin (PE) (Southern Biotechnologies, Birmingham, AL).

Techniques: Double Staining, Confocal Laser Scanning Microscopy, Staining, Derivative Assay

Journal:

Article Title: Bone-Marrow-Derived Cells Contribute to Glomerular Endothelial Repair in Experimental Glomerulonephritis

doi:

Figure Lengend Snippet: Strain-Specific MHC-I Expression on Peripheral Blood Leukocytes of WR, BN, and WR BM → BN Rats

Article Snippet: The following primary antibodies were used: fluorescein isothiocyanate (FITC)-labeled anti-BN-MHC-I (CL027F; Cedarlane, Hornby, Ontario, Canada), anti-WR-MHC-I (U9F4 19 ; kindly provided by J. Rozing, Groningen, the Netherlands), and secondary antibody goat anti-mouse-phycoerythrin (PE) (Southern Biotechnologies, Birmingham, AL).

Techniques: Expressing

Journal: Cancer Microenvironment

Article Title: Participation of Tumor-Associated Myeloid Cells in Progression of Amelanotic Melanoma (RMM Tumor Line) in F344 Rats, with Particular Reference to MHC Class II- and CD163-Expressing Cells

doi: 10.1007/s12307-017-0193-x

Figure Lengend Snippet: Probes used in the real-time RT-PCR

Article Snippet: The data were analyzed using the comparative C t method ( ΔΔ C t method). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Symbol Name Assay ID RT1-Ba RT1 class II, locus Ba Rn01428452_m1 IFN-γ Interferon gamma Rn00594078_m1 M-CSF Colony stimulating factor-1 Rn00572010_m1 GM-CSF Colony stimulating factor-2 Rn01456850_m1 TNF-α Tumor necrosis factor-alpha Rn01525859_g1 TGF-β1 Transforming growth factor-beta 1 Rn00572010_m1 Il-1β Interleukin 1 beta Rn00580432_m1 IL-6 Interleukin-6 Rn01410330_m1 IL-10 Interleukin-10 Rn00563409_m1 IL-12a Interleukin-12a Rn00584538_m1 CCL2 Chemokine (C-C motif) ligand 2 Rn00580555_m1 CCL5 Chemokine (C-C motif) ligand 5 Rn00579590_m1 CXCL1 C-X-C motif chemokine ligand 1 Rn00578225_m1 ΗΙΦ−1α Hypoxia inducible factor-1 alpha Rn01472831_m1 VEGFa Vascular endothelial growth factor A Rn01511602_m1 Flt-1 FMS-related tyrosine kinase 1 Rn01409533_m1 18 s rRNA Eukaryotic 18S rRNA Hs99999901_s1 Open in a separate window Probes used in the real-time RT-PCR table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene name Sequence Galectin-3 (forward) 5′-ATCCTGCTACTGGCCCTTT-3′ (reverse) 5′-GCGATGTCGTTCCCTTTCTT-3′ MMP-2 (forward) 5’GGGTCTATTCTGCCAGCACTTT-3′ (reverse) 5’CGGGGTCCATTTTCTTCTTTACTT-3′ MMP-9 (forward) 5’TGCCCTGGAACTCACACAAC-3′ (reverse) 5’GGAAACTCACACGCCAGAAG-3’ TIMP-1 (forward) 5’CCTCTGGCATCCTCTTGTTG-3′ (reverse) 5’GGTGGTCTCGATGATTTCTGG-3’ TIMP-2 (forward) 5’CGTTTTGCAATGCAGACGTA-3′ (reverse) 5’GGCCGTGTAGATAAATTCGATG-3’ PDGFb (forward) 5’TCTGGCCTGCAAGTGTGAG-3′ (reverse) 5’CCCGAGTTTGAGGTGTCTTG-3’ 18 s rRNA (forward) 5’-GTAACCCGTTGAACCCCATT-3′ (reverse) 5′-CCATCCAATCGGTAGTAGCG-3’ Open in a separate window Primers used in the real-time RT-PCR Cell Counts and Statistics Serial sections were used for cell counting.

Techniques:

Journal: Cancer Microenvironment

Article Title: Participation of Tumor-Associated Myeloid Cells in Progression of Amelanotic Melanoma (RMM Tumor Line) in F344 Rats, with Particular Reference to MHC Class II- and CD163-Expressing Cells

doi: 10.1007/s12307-017-0193-x

Figure Lengend Snippet: Laser microdissection (LMD) and real-time RT-PCR for MHC II-HIGH and MHC II-LOW regions in RMM nodules (0.5 cm and 3 cm). (a) example of MHC II-HIGH and MHC-LOW areas detected by immunofluorescence staining with Alexa 488-conjugated anti-MHC class II, before microdissection. Real-time RT-PCR for RT1-Ba (b), IFN-γ (c), GM-CSF (d), IL-12a (e), M-CSF (f), TGF-β1 (g), IL-10 (h) and HIF-1α (i). Tukey’s test *P < 0.05, **P < 0.001, significantly different from MHC II-LOW within each size point; †P < 0.05, ††P < 0.001, significantly different from MHC II-HIGH in 0.5 cm tumors. Bar, 50 μm

Article Snippet: The data were analyzed using the comparative C t method ( ΔΔ C t method). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Symbol Name Assay ID RT1-Ba RT1 class II, locus Ba Rn01428452_m1 IFN-γ Interferon gamma Rn00594078_m1 M-CSF Colony stimulating factor-1 Rn00572010_m1 GM-CSF Colony stimulating factor-2 Rn01456850_m1 TNF-α Tumor necrosis factor-alpha Rn01525859_g1 TGF-β1 Transforming growth factor-beta 1 Rn00572010_m1 Il-1β Interleukin 1 beta Rn00580432_m1 IL-6 Interleukin-6 Rn01410330_m1 IL-10 Interleukin-10 Rn00563409_m1 IL-12a Interleukin-12a Rn00584538_m1 CCL2 Chemokine (C-C motif) ligand 2 Rn00580555_m1 CCL5 Chemokine (C-C motif) ligand 5 Rn00579590_m1 CXCL1 C-X-C motif chemokine ligand 1 Rn00578225_m1 ΗΙΦ−1α Hypoxia inducible factor-1 alpha Rn01472831_m1 VEGFa Vascular endothelial growth factor A Rn01511602_m1 Flt-1 FMS-related tyrosine kinase 1 Rn01409533_m1 18 s rRNA Eukaryotic 18S rRNA Hs99999901_s1 Open in a separate window Probes used in the real-time RT-PCR table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene name Sequence Galectin-3 (forward) 5′-ATCCTGCTACTGGCCCTTT-3′ (reverse) 5′-GCGATGTCGTTCCCTTTCTT-3′ MMP-2 (forward) 5’GGGTCTATTCTGCCAGCACTTT-3′ (reverse) 5’CGGGGTCCATTTTCTTCTTTACTT-3′ MMP-9 (forward) 5’TGCCCTGGAACTCACACAAC-3′ (reverse) 5’GGAAACTCACACGCCAGAAG-3’ TIMP-1 (forward) 5’CCTCTGGCATCCTCTTGTTG-3′ (reverse) 5’GGTGGTCTCGATGATTTCTGG-3’ TIMP-2 (forward) 5’CGTTTTGCAATGCAGACGTA-3′ (reverse) 5’GGCCGTGTAGATAAATTCGATG-3’ PDGFb (forward) 5’TCTGGCCTGCAAGTGTGAG-3′ (reverse) 5’CCCGAGTTTGAGGTGTCTTG-3’ 18 s rRNA (forward) 5’-GTAACCCGTTGAACCCCATT-3′ (reverse) 5′-CCATCCAATCGGTAGTAGCG-3’ Open in a separate window Primers used in the real-time RT-PCR Cell Counts and Statistics Serial sections were used for cell counting.

Techniques: Laser Capture Microdissection, Quantitative RT-PCR, Immunofluorescence, Staining