Figure 3. Generation and characterization of GECs with or without Cd151.

(A) GECs isolated from an adult Cd151^fl/fl;Trp53^+/– mouse. (B) Cell surface expression of Cd151 was absent after Cre-mediated recombination in vitro, as shown by FACS. GEC+, Cd151-positive GECs; GEC-, Cd151-negative GECs. (C) Immunofluorescent analysis of confluent Cd151-positive and -negative GEC monolayers showing similar cell size as well as comparable actin, nephrin, podocin, and cadherin distribution. (D) Western blot analysis of whole cell lysates (WCL) showing absence of Cd151 in the Cd151-negative GECs and CD151 being coimmunoprecipitated with α3 in Cd151-positive, but not Cd151-negative, GECs. (E) Western blot analysis of immunoprecipitations of α3 and FLAG-tagged human CD151 rescue constructs. CD151 coimmunoprecipitated with α3 only when the QRD integrin-binding motif was intact. GEC-re^WT, Cd151-negative GECs rescued with human wild-type CD151; GEC-re^QRD*, Cd151-negative GECs rescued with the mutated ¹⁹⁴QRD–INF¹⁹⁶ sequence of human CD151. Scale bars: 100 μm (A, top, and C, triple stain), 25 μm (A, bottom, and C, single stains).