Figure 3.

DENND5A Is Detected Predominantly in Developing Neurons

(A) Equal protein aliquots of lysates from various tissues were blotted with antibodies recognizing DENND5A (polyclonal antibody, Abcam) and actin (monoclonal antibody C4, Millipore) as indicated. Lysates from HEK293T cells expressing FLAG-tagged DENND5A (FLAG-DENND5A) were included as a control.

(B) Crude lysates were prepared from brains dissected from embryonic day 18 (E18) or postnatal day 5, 30, or 60 (P5, P30, or P60) rats. Equal protein aliquots were processed for western blot with antibodies recognizing DENND5A and GAPDH (monoclonal antibody, Santa Cruz) as indicated.

(C) Primary rat cortical neurons were seeded in culture dishes, and lysates were prepared from the neurons at the indicated days in vitro. Equal protein aliquots were processed for western blots with antibodies recognizing DENND5A, GAPDH, and synapsin.¹⁹

(D) Quantification of the relative amounts of DENND5A and synapsin from blots as in (C). The highest-intensity values were set as 1.0. Data represent the mean ± SEM from eight independent experiments.