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. Author manuscript; available in PMC: 2016 Mar 10.
Published in final edited form as: Cancer Lett. 2014 Nov 20;357(1):231–241. doi: 10.1016/j.canlet.2014.11.036

Fig. 3.

Fig. 3

Expression and tyrosine phosphorylation status of HSC70 in L1210/0 and L1210/DDP cells. (A) HSC70 expression in different cellular fractions including the plasma membrane and cytosol fractions in both L1210/0 (S) and L1210/DDP (R) cells by Western blotting assay. Beta-actin and Na-K-ATPase were used as a control to confirm the purity of the different cellular fractions. The blots shown are representative of three independent experiments. (B) Comparison of the tyrosine phosphorylation status in the membrane fraction between L1210/0 and L1210/DDP cells by Western blotting assays used PY69 antibody. A tyrosine-phosphorylated band around 70 kDa was observed in the membrane fraction of L1210/0 cells. This band was not found in the L1210/DDP cells. The blots shown are representative of three independent experiments. (C) HSC70 expression in the membrane fraction in both sensitive L210/0 and resistant L1210/DDP cells by Western blotting and immunoprecipitation assays. The membrane fractions from both L1210/0 and L1210/DDP cell lines were immunoprecipitated by PY20 antibody agarose beads, followed by a Western blotting assay for the detection of HSC70 in the immunoprecipitated mixture. HSC70 was detected in the membrane fraction after it was immunoprecipitated with PY20 antibody agarose beads from L1210/0 cells. However it was hardly detected in the membrane fraction from L1210/DDP cells. The blots shown are a representation of three independent experiments.