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. 2012 Oct 8;7(10):e39723. doi: 10.1371/journal.pone.0039723

Figure 2. Down-regulation of Mybbp1a in primary wild type MEFs and in NIH3T3 cells has opposite effects on cell proliferation.

Figure 2

(A) Down regulation of Mybbp1a in MEFs with the specific lentiviral vector shRNA3 induces early senescence, as opposed to a non-target empty (NT) vector. (B) Immunoblot of the cells of panel A showing down regulation of MYBBP1A (p160), using tubulin as control. (C) Growth rate determination (crystal violet assay, see Methods) in Mybbp1a down-regulated NIH3T3 cells with lentiviral vectors expressing specific Sh1RNA or Sh3RNA, as opposed with control empty Non Target vector (NT). Crystal violet assays were performed over 10 days counting the cells in triplicate every 2 days. The p-value of the difference between Sh3 and NT-treated cells was ≤0.001 (t test). (D) Immunoblot of the cells of panel C. (E) Growth rate determination of MYBBP1A down-regulated HeLa cells by specific siRNAs (see Methods). Ctl: untreated cells. Lipo: transfection control with lipofectamine only. High GC and Medium GC: two control oligonucleotides (indicated by the siRNAs manufacturer) of high and, respectively, medium GC content. siRNA1, siRNA2 and siRNA3: specific MYBBP1A siRNAs (see Methods for sequences). (F) Immunoblots of the cells of panel B upon culturing for 24, 48, 72 and 96 h after transfection.