Supplementary Figure 1: Specificity test of custom polyclonal antibodies used in this study.
From: Differential centrifugation–based biochemical fractionation of the Drosophila adult CNS
Western blot analysis of adult fly head extracts probed with affinity-purified (a) anti-DRBPSH3 II+III and (b) anti-DLiprin-αC-term antibodies. (a) RBP expression analysis in control (rbp heterozygous animals: S201/+) and rbp mutant lines (rbpSTOP3/S201 and rbpMinos/S201). Two exposures of the same blot to light-sensitive autoradiography films are presented: 10 seconds (upper panel) and 30 seconds (middle panel). Bands of approximately 160 kDa and 190 kDa are detected (corresponding to the predicted sizes of RBP isoforms) in control animals only. An unspecific signal at around 130 kDa is also observed (most likely due to cross-reactivity of the anti-DRBPSH3 II+III antibody with other proteins in the fly head extract). (b) Liprin-α expression analysis in the indicated wild-type control (+/+ = w1118), heterozygous liprin-α animals (lipF3ex15/+) and liprin-α mutants (lipF3ex15/lipEPexR60). A single band around 140 kDa, corresponding to the predicted size of Liprin-α, is observed in control animals and is reduced in liprin-α heterozygous animals, with only faint signal in the hypomorphic liprin-α mutant lipF3ex15/lipEPexR60. α-Tubulin is displayed as a loading control for each western blot.
