Journal: Molecular cancer research : MCR

Article Title: Suppression of AKT Phosphorylation Restores Rapamycin-based Synthetic Lethality in SMAD4-defective Pancreatic Cancer Cells

doi: 10.1158/1541-7786.MCR-12-0679

Figure Lengend Snippet: Knockdown of eIF4E is sufficient to induce apoptosis in Smad4-null pancreatic cancer cells in the presence of serum. A, Panc1 cells were plated as in Fig. 1 and then shifted to media containing 10% serum for 24 hours. Cells were then transfected with either control, Smad4, eIF4E, or dual Smad4-eIF4E siRNA. Forty-eight hours later, the cells were harvested and analyzed by Western blot analysis for levels of cleaved PARP, Smad4, and eIF4E. B, BxPC3 cells were plated as above then transfected with either control or eIF4E siRNA. Cells were harvested 48 hours later and analyzed by Western blot analysis for levels of cleaved PARP and eIF4E. Western blot analyses are representative of at least 2 independent experiments.

Article Snippet: Smad4 and non-targeted negative control siRNA duplexes were obtained from Santa Cruz Biotechnology.

Techniques: Knockdown, Transfection, Control, Western Blot

Journal: EMBO Reports

Article Title: G‐Quadruplexes act as sequence‐dependent protein chaperones

doi: 10.15252/embr.201949735

Figure Lengend Snippet: A A schematic illustration of expression vectors. Both the expression of protein folding enhancing factors and TagRFP675 are under the control of pBAD promoter, which is induced by 0.2% L ‐Arabinose. B Harvested cells. Empty vector (negative control), molecular chaperones (GroEL, DnaK, Hsp33, Spy, ClpA, IbpA, and IbpB), and selected RNA sequences (Seq42, Seq359, Seq536, and Seq576) were used as protein folding enhancers. NI and IN indicate: non‐induced and induced, respectively. C Cellular fluorescence assay of TagRFP675 with various protein folding enhancing factors. Protein expression was induced at 42°C, and the fluorescence of each sample was measured with spectrophotometer. White bars (+Empty IN and + Seq42) indicate negative controls. The data are shown as mean ± SD of technical triplicates ( n = 3).

Article Snippet: For the expression of biosensors, we used pBAD/HisD‐TagRFP675 and pBAD18‐wtGFP. pBAD/HisD‐TagRFP675 was a gift from Vladislav Verkhusha (Addgene plasmid # 44274; http://n2t.net/addgene:44274 ; RRID:Addgene_44274) (Piatkevich et al , ). pBAD18‐wtGFP was given by Jonathan S. Weismann’ laboratory (Wang et al , ).

Techniques: Expressing, Plasmid Preparation, Negative Control, Fluorescence, Spectrophotometry