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. 2019 Aug 6;10(47):4868–4879. doi: 10.18632/oncotarget.27115

Figure 2. AUF1 positively controls the expression of VEGF-A.

Figure 2

(A and B) U2OS and HOS cells were transfected with specific AUF1siRNA (3 different sequences) or pSILENCER- AUF1siRNA and scrambled sequences were used as controls. The generated cells (AUF1si-A, AUF1si-B, AUF1si-C and pSILENCER-AUF1siRNA) as well as their respective controls were used to prepare total RNA, which was then utilized to assess the levels of the VEGF-A and AUF1 mRNAs by qRT-PCR. Error bars represents means ± SD. ** p < 0.001. (C) Cell lysates were prepared from the indicated cells and were used for immunoblotting utilizing specific antibodies. (C) SFCM were collected from the indicated cells after 24 hrs of culture and the level of the secreted VEGF-A was determined by ELISA. Error bars represent means ± SD of 3 different experiments. *** p < 0.00002. (D) p37AUF1 isoform was ectopically expressed in SaOS2 cells using empty vector as control. The generated cells (Control and p37AUF1) were utilized to prepare total RNA and the level of the indicated transcripts were assessed by qRT-PCR using specific primers. Error bars represent means ± SD. (E) whole cell lysates were prepared from the indicated cells and the levels of the indicated proteins were assessed by immunoblotting using specific antibodies. (F) SFCM were collected from the indicated cells after 24 hrs of culture and the level of the secreted VEGF-A was determined by ELISA. Error bars represent means ± SD of 3 different experiments. *** p < 0.0000321.