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. Author manuscript; available in PMC: 2017 May 1.
Published in final edited form as: Cell Signal. 2016 Feb 11;28(5):448–459. doi: 10.1016/j.cellsig.2016.02.005

Figure 5. Shoc2 silencing results in reduced expression and secretion of LGALS3BP.

Figure 5

(A) The expression levels of LGALS3BP were quantified in Cos1, T47D and MCF7 cells depleted of Shoc2 using RT-PCR analysis and results were compared to those obtained by the RNA-seq approach. Data are presented as the log2fold change of the Shoc2 mRNA levels in Shoc2-depleted cells normalized to control (NT) (mean ± SD; n = 3).

(B) Cos1, T47D, and MCF7 cells depleted of Shoc2 (LV1) and control cells (NT) were harvested for immunoblotting. The expression of LGALS3BP in culture medium and cell lysate was analyzed using specific antibodies. Other indicated proteins were analyzed using specific antibodies.

(C) Cos-NT and Cos-LV1 cells were transiently transfected with His-LGALS3BP. At 48h post-transfection, His-LGALS3BP was immunoprecipitated using anti-His antibodies and then digested by Endoglycosidase H or Peptide -N-Glycosidase F (Endo H or PNGase F). Immunoprecipitates were analyzed by immunoblotting using anti-LGALS3BP antibody.

(D) Cos1 cells depleted of KSR1 (KSR1#2) and control cells (NT and LV1) were harvested for immunoblotting. LGALS3BP in culture medium was analyzed using specific anti-LGALS3BP antibodies.