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. 2011 May 3;152(7):2678–2689. doi: 10.1210/en.2010-1477

Fig. 4.

Fig. 4.

Expression of JunB is not required for 3T3-L1 differentiation. 3T3-L1 cells were infected with the MISSION TRC lentiviral particles coding for JunB shRNA (two independent replicates) or a nontargeting shRNA as control (ntc), selected for puromycin resistance, expanded as a mixed population, and induced to differentiate using MDI cocktail. A, JunB silencing has no impact on the expression of genes important for differentiation and lipolysis in differentiating 3T3-L1 cells. Expression of C/EBPß, PPARγ, C/EBPα, ATGL, and HSL in JunB-silenced 3T3-L1 cells was compared with control cells (ntc) at d 3 and 6. B, Western blot analysis was performed to confirm silencing of JunB on protein level (two independently silenced 3T3-L1 cells named sil1 and sil2). β-Actin served as loading control. C, TG accumulation in 3T3-L1 cells is unchanged upon silencing of JunB (JunB_sil1 and JunB_sil2) in comparison with ntc. TG content of the cell lysates was measured on d 3, 6, and 10 after induction of differentiation and normalized to protein content. D, Oil red O staining of control and JunB-silenced 3T3-L1 cells on d 10 of differentiation.