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. Author manuscript; available in PMC: 2015 Oct 15.
Published in final edited form as: Exp Cell Res. 2014 Jul 9;328(1):99–117. doi: 10.1016/j.yexcr.2014.06.022

Figure 3.

Figure 3

miR-21 increases phosphorylation of Akt and its downstream effectors by targeting PDCD4. ACHN as well as 786-O renal cancer cells were transfected with miR-21 Sponge along with siRNAs against PDCD4. The cell lysates were immunoblotted with indicated phospho-specific antibodies and other antibodies as described in the legends of Fig. 2. In panels A and C, same membranes for each panel were immunoblotted with phospho-antibodies, total antibodies of the same protein and actin antibody as indicated. Same samples for each panel were run in parallel to immunoblot with PDCD4 antibody. For panel B, same blot was used to probe with phospho-IKKβ and PDCD4 antibodies. Same lysates were run in parallel to probe with IKKβ and actin antibodies. In panel D for ACHN cells, same blot was used to probe with phospho-p65 and p65 antibodies. Same lysates were run in parallel to probe with PDCD4 and actin antibodies. For 786-O cells, same blot was used to probe with p65, PDCD4 and actin antibodies. Same lysates were run in parallel to probe with phospho-p65. Quantifications of the results and are shown in Supplementary Fig. S5. Expression of miR-21 Sponge for these results was determined in parallel experiments and is shown in Supplementary Fig. S5.