Fig. 5. Immunofluorescent staining of CD9 on the surface of CD9^–/– eggs injected with either wild-type or SFQ(173-175)AAA mutant mouse CD9 mRNA. After in vitro fertilization, eggs were fixed and washed as described in Materials and Methods. The fixed eggs were incubated with anti-mouse CD9 antibody KMC8 as the primary antibody, followed by a goat anti-rat IgG (H+L) conjugated with Alexa Fluor^TM 488 as the secondary antibody. Immunofluorescent staining was obtained with a laser scanning confocal microscope (Carl Zeiss, LSM 410). (A) Immunofluorescent staining on CD9^–/– eggs injected with wild-type mouse CD9 mRNA. (B) Immunofluorescent staining on CD9^–/– eggs injected with SFQ-to-AAA mutated mouse CD9 mRNA. (C,D) Phase contrast images of A and B, respectively. Fluorescence levels were somewhat variable within each group of eggs injected with the identical amount and type of mRNA.

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