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. Author manuscript; available in PMC: 2015 Oct 15.
Published in final edited form as: Sci Transl Med. 2014 Oct 15;6(258):258ra140. doi: 10.1126/scitranslmed.3010157

Fig. 4.

Fig. 4

Blockade of PirB binding enhances OD plasticity in WT visual cortex. (A) Schematic of soluble PirB-Myc-His (sPirB) fusion protein, indicating extracellular Ig-like domains plus Myc and His tags. (B) Western blot of culture supernatant from sPirB-transfected HEK293 cells, detecting Myc tag and PirB ectodomain; Myc-His–tagged alkaline phosphatase (AP-Myc-His) is a positive control. (C) PirB phosphorylation is decreased after 7 days (P21 to P28) of sPirB infusion into WT mouse cortex, as shown by phospho-tyrosine IP and PirB Western blot of cortical lysates from infused (sPirB infused), uninfused (sPirB uninf.) hemispheres, or untreated littermate controls. (D) Section of visual cortex immunostained with anti-Myc antibody after 11 days (P21toP32)of sPirB or BSA infusion (1 mg/ml). Scale bar, 1 mm. (E and F) Minipump infusions of sPirB during critical period (CP). Timeline as shown. (E) Example Arc mRNA in situ hybridization micrographs of visual cortex after BSA (top)or sPirB (bottom) treatment. Scale bar, 500 mm. Red asterisks indicate borders of Arc mRNA signal induced by stimulating the ipsilateral (open) eye in layer 4. (F) Graphs comparing width of Arc mRNA signal in L4 after open-eye stimulation. Width of territory activated by open-eye stimulation after ME is greater after sPirB infusion than with BSA. NRBSA: n = 4 mice, NR sPirB: n = 4, MEBSA: n = 5 versus ME sPirB: n = 6, P < 0.0001, by two-way ANOVA and Tukey post hoc test for all comparisons indicated. (Gto I) sPirB infusions into adult WT visual cortex; timeline as shown. (G) Example of Arc mRNA situ hybridization micrographs at P74. Scale bar, 500 mm. (H) Graphs comparing width of Arc mRNA signal in L4 after stimulation of the ipsilateral (open) eye. sPirB infusion from P63 to P74 enhances open-eye expansion after ME. NRBSA: n = 4 mice versus NR sPirB: n = 4, P = 0.99. MEBSA: n = 4 versus ME sPirB: n = 5, P = 0.0004. NR versus MEBSA: P = 0.88, NR versus ME sPirB: P < 0.0001. (I) sPirB infusion coupled with 3 days of MD also enhances OD plasticity. (J) sPirB infusion has no effect on OD plasticity when infused into visual cortex of PirB−/− mice. ME PirB−/− BSA: n = 5 mice versus ME PirB−/− sPirB: n = 5, P = 0.95, ME PirB−/− BSA versus MEWTBSA: P = 0.034. MD BSA versus MD sPirB: n = 4 mice per group, P = 0.036. *P < 0.05, ***P < 0.001, ****P < 0.0001, by two-way ANOVA and Tukey post hoc test.