1. Proteasome Inhibition Sensitizes Non–Small-Cell Lung Cancer to Gemcitabine-Induced Apoptosis 
Chadrick E. Denlinger, MD, Brian K. Rundall, DO, Michael D. Keller, BS, David R. Jones, MD 
The Annals of Thoracic Surgery 
Volume 78, Issue 4, Pages (October 2004)
DOI: /j.athoracsur

2. Fig 1
Non–small-cell lung cancer cells were treated with escalating doses of gemcitabine (1 to 200 μmol/L) for 24 hours. Cell survival was determined by staining with crystal violet dye.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )

3. Fig 2
(A) Nuclear extracts from non–small-cell lung cancer (NSCLC) cells treated with nothing, gemcitabine (Gem; 10 μmol/L), bortezomib (Velcade; Vel; 50 nmol/L), or both drugs were analyzed by Western blot analysis probing for the transcriptionally active nuclear factor (NF)-κB subunit p65. Blots were reprobed for RNA pol II as a loading control. (B) DNA binding of NF-κB was determined in the A549 cell line after identical treatment conditions by electrophoretic mobility shift assay. (C) Messenger RNAs from similarly treated NSCLC cells were evaluated by reverse transcription-polymerase chain reaction to determine c-IAP2, Bcl-xL, and interleukin (IL)-8 expression. (GAPDH = glyceraldehyde phosphate dehydrogenase; TNF = tumor necrosis factor.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )

4. Fig 3
(A) Non–small-cell lung cancer cells were treated for 12 hours with nothing, gemcitabine (Gem; 10 μmol/L), bortezomib (Velcade; Vel; 50 nmol/L), or both drugs. Western blot analysis was performed to probe for p21 and p53. Blots were reprobed for β-tubulin as a loading control. (B) Cell-cycle profiles were determined by fluorescence-activated cell sorting analysis in the A549 cells after 12 hours of treatment.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )

5. Fig 4
Apoptosis was determined in non–small-cell lung cancer cells after 24 hours of treatment with nothing, gemcitabine (gem; 10 μmol/L), bortezomib (Velcade; 50 nmol/L), or both drugs by quantifying (A) caspase-3 activity and (B) DNA fragmentation (‡p ≤ 0.004; †p ≤ 0.003; *p = 0.02). ▧ = no treatment; ▨ = Velcade; = gemcitabine; ■ = gem/Velcade. (RFU = relative fluorescence units; OD = optical density.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )

6. Fig 5
A549 non–small-cell lung cancer xenografts were developed in athymic nude mice. Treatment was initiated when tumors achieved a volume of 0.5 cm3. Tumors were measured and volumes were calculated every other day over a 4-week treatment period (p ≤ 0.04 versus gemcitabine). (Gem = gemcitabine; Vel = Velcade [bortezomib].)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )