FIGURE 7.

Biochemical interactions between Ringer and Futsch. (A–G) Representative immunoblots from larval lysates (A–C) and fly head lysates (D–G) showing total levels of Ringer (A,D), Futsch (B,E), Ac-Tub (C,G), and Tub (F) of specified genotypes. Actin (A–G) was used as the loading control. (H) Quantification of the ratio of band intensities of Ac-Tub/Actin in represented genotypes. Adult head lysates of wild type (+/+) and ringer–/– probed with anti-Ringer (I); wild type and futsch–/– probed with anti-Futsch (J) and wild type probed with anti-Tub (K) antibodies. Head lysates of wild type in combination with ringer–/– (I) and futsch–/– (J) immunoprecipitated with anti-Ringer and anti-Futsch, respectively. There is presence of a non-specific band (asterisk, I) in wild type and mutant lysate and IP lanes when probed with rat anti-Ringer antibodies. Wild type head lysates immunoprecipitated with anti-Tub (K). (L–N) Head lysates of wild type (+/+) in combination with ringer–/– (L) and futsch–/– (M) co-immunoprecipitated with anti-Tub and anti-Futsch, respectively and probed with anti-Ringer (L) and anti-Tub (M). Head lysates from elav-Gal4;UAS-m-Cherry-Ringer, and ringer–/– probed with anti-m-Cherry (N) and co-immunoprecipitation of lysates from elav-Gal4;UAS-m-Cherry-Ringer, and ringer–/– probed with anti-m-Cherry (N). (O–Q) Adult wild type head lysates probed with anti-Ringer (O) and anti-Futsch (Q). Immunoblots of GST-Control and GST-Ringer probed with anti-Ringer (P) and anti-Futsch (Q). Note that blots separated by a space (N,Q) were probed separately. futsch^K68/K68 mutants were used for all biochemical analyses. n = 3 (each experiment was done independently three times and the most representative blots are shown).