Localization of Smad4 and pSmad1/5/8 on early endosomes in E7 chicken lenses

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> A. EEA1 (green), Smad4 (red) and TOPRO (blue). C. EEA1 (green), pSmad1/5/8 (red) and TOPRO (blue). B and D are higher magnification images of the regions outlined in figures A and C respectively. The insets in B and D show 2X higher magnification images of antibody-stained cytoplasmic vesicles. Both Smad4 and pSmad1/5/8 show co-localization (yellow) with EEA1

Double staining of lens sections with antibody against EEA1 (green) and phalloidin (red) in A and antibody to EEA1 (green) and antibody to β-tubulin (red) in B

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> No co-localization of either filamentous actin or tubulin with EEA1 was detected (LE- lens epithelium, LF- lens fibers)

Marked reduction in the endosomal and nuclear localization of pSmad1 in lenses

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> A. WT (Cre-negative) lens; Rab5B (green), pSmad1 (red). B. (Cre-positive) lens Rab5B (green), pSmad1 (red). Both the number of endosomes and the relative number of Smad1-positive endosomes is reduced in the lens. In B, note the relative difference in staining intensity for pSmad1 and Rab5b in the ciliary epithelium (CE), compared to the lens

Localization of pSmad1 and pSmad2 on late endosomes in P3 mouse lens cells

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> A. Rab7 (red), pSmad1 (green), TOTO-1 (blue). B. Rab7 (red), pSmad2 (green), TOTO-1 (blue). Both pSmad1 and pSmad2 co-localize with Rab7 on a small number of cytoplasmic vesicles. Nuclear staining for pSmad1 and pSmad2 is mostly obscured by the strong fluorescence of the nucleic acid stain, TOTO-1

Endosomal localization of pSmad1, pSmad2, TGIF, C184M and c-Ski in P3 mouse lenses

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> A. Rab5B (red), pSmad1 (green). B. EEA1 (green), pSmad2 (red). C. EEA1 (green), TGIF (red). D. EEA1 (green), C184M (red). E. c-Ski (green), C184M (red). F is a diagram of the neonatal mouse lens showing the regions that are represented in each of the images

Double immunostaining of lens sections with two rabbit primary antibodies (A, C) and controls in which the second primary antibody was omitted (B, D)

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> A. Rab5B (red), pSmad1 (green). B. Rab5B (red), no pSmad1 primary antibody. No green staining from the second anti-rabbit secondary antibody was detected. C. Rab7 (green), Rab5B (red), TOTO-1 (blue). D. Rab7 (green), no Rab5B primary antibody, TOTO-1 (blue). No red staining from the second anti-rabbit secondary antibody was detected. E. Diagram illustrating the method used for double labeling () and the alternative outcomes of the control studies for specificity (). In , the first rabbit primary antibody (light blue, 1) binds to its antigen and is localized by a fluorescent-labeled anti-rabbit secondary antibody (red, 2). The tissue is washed, fixed in formalin and washed again. The second rabbit primary antibody is then added (yellow), which binds to the second antigen (3). The second anti-rabbit secondary antibody (green) is added and binds to the second rabbit primary antibody (4). The sequence shown in and illustrates the controls used in the present study. The first two steps are as in , but the second rabbit primary antibody is omitted (3), providing no antibody for the second anti-rabbit fluorescent antibody to bind (green, 4). In , the second fluorescent-labeled anti-rabbit secondary antibody (green) does not bind the first primary antibody (blue) and the section remains singly labeled. In the steps shown in , the second fluorescent-labeled anti-rabbit secondary antibody (green) binds to the first rabbit primary antibody (blue), resulting in spurious "co-localization." Since staining by the second anti-rabbit secondary antibody was not seen, as shown in B and D, accurately reflects the results obtained in the present studies; the events shown in were not observed

Localization of Smad4 (A), Smad7 (B),) and Smad6 (C), c-Ski (D) and TGIF (E) in lens epithelial (LE) and fiber cells (LF)

<p><b>Copyright information:</b></p><p>Taken from "Intracellular mediators of transforming growth factor β superfamily signaling localize to endosomes in chicken embryo and mouse lenses in vivo"</p><p>http://www.biomedcentral.com/1471-2121/8/25</p><p>BMC Cell Biology 2007;8():25-25.</p><p>Published online 25 Jun 2007</p><p>PMCID:PMC1914053.</p><p></p> Nuclear localization of Smad4 and TGIF is seen mainly in lens fiber cells. Smad7 is seen in the nuclei of both epithelial and fiber cells, although staining is stronger in fiber cell nuclei. Both I-Smads, Smad7 and Smad6, are abundant in the cytoplasm of lens epithelial cells. c-Ski is found mainly to the cytoplasm. A-C are sections of E7 chicken lenses and D-E are P3 mouse lenses

Weight-of-Evidence Analysis for the Development of a Reference List of Chemical Respiratory Sensitizers

Poster for SOT on March 19-23, 2023 in Nashville TN Science Inventory, CCTE products: https://cfpub.epa.gov/si/si_public_search_results.cfm?advSearch=true&showCriteria=2&keyword=CCTE&TIMSType=&TIMSSubTypeID=&epaNumber=&ombCat=Any&dateBeginPublishedPresented=07/01/2017&dateEndPublishedPresented=&dateBeginUpdated=&dateEndUpdated=&DEID=&personName=&personID=&role=Any&journalName=&journalID=&publisherName=&publisherID=&sortBy=pubDate&count=25</p