LACK expression is maintained under serum depletion.
(A) Evaluation of relative abundance of the LACK antigen by Western blot performed with the anti-LACK polyclonal antibody (diluted 1:500) over total protein extracts of L. infantum promastigotes grown under HIFBS-depletion or in CM. The anti-gGAPDH polyclonal antibody (diluted 1:10,000) was used as the loading control antibody. The secondary antibody was HRP-conjugated goat anti-rabbit IgG (diluted 1:2,000). Chemoluminescence detection was performed with the ECL reagents (GE Healthcare) and the membrane was developed in an X-ray film. Upper bands are likely aggregates of the LACK protein . (B) Bar graph of the relative expression ratio of the LACK antigen with respect to the gGAPDH. Mean and SD of three biological replicates are represented. Densitometry was performed with Gel Doc XR System and Quantity One version 4.6. software (BioRad). Both groups were compared by the Student's t-test including three biological replicates.