Anti RFP  

Western blot of RFP recombinant protein detected with Rockland's unconjugated polyclonal anti-RFP antibody (600-401-379). Lane 1 shows no reaction against a GFP recombinant protein present in 10 ug of HeLa cell extract. Lane 2 shows a single band detected in 10 ug of a HeLa lysate containing RFP recombinant protein. Rockland's polyclonal anti-RFP detects a 27 kDa band corresponding to the epitope tag RFP. A 4-12% Bis-Tris gradient gel (Invitrogen) was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the unconjugated primary antibody diluted 1:2,500 for 1 h at room temperature followed by washes and reaction with a 1:5,000 dilution of IRDye^TM800 conjugated Goat-a-Rabbit IgG [H&L] MX (611-132-122). IRDye^TM800 fluorescence image was captured using the Odyssey^(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.

(B) Example AAV-EGFP-labeled callosal axon projection into contralateral V1/V2L border region where ChCs express RFP. The region that received callosal input is labeled as callosal binocular zone (cBZ). Fig 1. PMID:？33290732

j, Representative images of tdTomato+/Sma+？cells in fibrous caps (bracketed in yellow dashed lines; quantified in？k) from indicated groups. Fig 5. PMID:？34746803

？(C) Co-immunostaining of RFP, RUNX1, CDH1 in the proximal AP. Arrows indicate RFP labeled RUNX1+？cells. Scale bar: 50 μm.？ Fig 4. ？PMID:？33025905

(E, F) Co-immunostaining of RFP, NKX3.1, CDH1 in UGS explants harvested at day 2 (E) and day 7 (F). Higher magnification images of (i) proximal and (ii) distal regions are shown for day 7. Arrows show RFP+？NKX3.1+？cells, arrowheads show RFP+？NKX3.1-？cells. Scale bars: 200 μm (yellow) and 50 μm (white). (H, I) Co-immunostaining of RFP, K4, CDH1 in UGS explants harvested at day 2 (H) and day 7 (I). Higher magnification images of (i) proximal and (ii) distal regions are shown for day 7. Arrows show RFP+？K4 cells, arrowheads show RFP+？K4+？cells. Scale bars: 200 μm (yellow) and 50 μm (white). Fig 7. ？PMID:？33025905

(A, B) Co-immunostaining of RFP, p63, CDH1 in UGS explants harvested at day 2 (A) and day 7 (B). Higher magnification images of (i) proximal and (ii) distal regions are shown for day 7. Arrows show RFP+？p63+？cells, arrowheads show RFP+？p63-？cells. Scale bars: 200 μm (yellow) and 50 μm (white). (C) Quantification of the percentage of epithelial RFP+？p63+？cells at day 2 (n？=？7) and day 7 (n？=？4) of UGS explant cultures. Quantification was performed within the boundaries delimited in？B？by dotted lines. (D) Co-immunostaining of RFP, K5, K8 in UGS explants harvested at day 2 (top) and day 7 (bottom). Higher magnification images of proximal (i) and (ii) distal regions are shown for day 7. Arrows show RFP+？K5+？K8+？cells, chevron arrows show RFP+？K5-？K8+, arrowheads show RFP+？K5+？K8-？cells. Scale bars: 200 μm (yellow) and 50 μm (white). (E) Quantification of the percentage of epithelial K5+？K8+, K5-？K8+？cells and K5+？K8-？cells in the RFP subset at day 2 (n？=？4) and day 7 (n？=？3) of UGS explant cultures. Quantifications were performed within the boundaries delimited in？D？by dotted lines. (F, G) Co-immunostaining of RFP,？Nupr1？(mRNA), CDH1 in UGS explants harvested at day 2 (F) and day 7 (G). Higher magnification images of (i) proximal and (ii) distal regions are shown for day 7. Arrows show RFP+？Nupr1+？cells, arrowheads show RFP+？Nupr1-？cells. Scale bars: 200 μm (yellow) and 50 μm (white). (J) Quantification of the percentage of epithelial K4+？cells in the RFP subset at day 2 (n？=？3) and day 7 (n？=？4) of UGS explant cultures. Quantification was performed within the boundaries delimited in？G？by dotted lines. Source files are available in？Figure 7―figure supplement 1―source data 1. ？PMID:？33025905