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. 2014 Mar 26;5(19):8959–8969. doi: 10.18632/oncotarget.1863

Figure 4. SPRY4-IT1 Knockdown Increases Lipin 2 Protein Accumulation in Melanoma Cells.

Figure 4

A) Affinity purification of SPRY4-IT1 from A375 cell lysates with SPRY4-IT1-specific probes followed by qPCR. SPRY4-IT1 is enriched compared to scrambled (control) probes. U1 RNA was used as endogenous control for pull-downs. (B) qPCR validation showing enrichment of SPRY4-IT1 following immunoprecipitation of A375 cell lysates with lipin 2-specific or control IgG antibodies. (C) Relative expression of SPRY4-IT1 in lipin 2 knock-down cells (D) Relative expression of lipin 2 after siRNA-mediated knockdown of SPRY4-IT1, SPRY4 exon 1 or lipin 2. (E) Phosphatidic acid phosphatase assay. (F) Western blot analysis of lipin 2 following siRNA-mediated knockdown of SPRY4-IT1 or lipin 2. All experiments were performed in triplicate.