Fig. 8. Depleting T6BP or NDP52 by RNAi KD increases the rate of constitutive secretion in HeLa cells. (A) HeLa cells expressing SEAP were either mock-transfected or transfected with siRNAs specifically targeting myosin VI, T6BP or NDP52 and were analysed by immunoblotting using anti-actin antibodies as a loading control. Myosin VI, T6BP and NDP52 expression was <10% of normal levels. (B) In HeLa cells depletion of T6BP or NDP52 or both proteins by single siRNA (P1) or SMARTpool (SP) primers causes a significant increase in SEAP secretion (30-90% increase), whereas depletion of myosin VI leads to a 80% decrease in secretion. The amount of SEAP activity in the growth medium was analysed by chemiluminescence and normalised relative to cell density.