Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: A) Expression of Stat3C-Flag in the whole lung of CCSP-rtTA/(tetO)7-CMV-Stat3C double-transgenic mice. Actin was used as control. WT: wild type mouse lungs; -Dox: doxycycline untreated bouble transgenic mouse lungs; +Dox1 & +Dox2: doxycycline treated mouse lungs. B) Expression of CHI3L1 protein in BALF of CCSP-rtTA/(TetO) 7 -CMV-Stat3C double-transgenic mice. Lane 1–3, doxycycline-untreated mice (Dox-); Lane 4–6, doxycycline-treated mice without showing lung tumor (Dox+); Lane 7–10, doxycycline-treated mice showing lung tumor (Lung Cancer).

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques: Expressing, Transgenic Assay

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: Left: CHI3L1 protein concentrations in BALF and serum of doxycycline-treated and untreated bitransgenic mice. Right: ROC (Receiver Operating Characteristic) curve analyses to determine the area under the curve (AUC). Dox-: doxycycline-untreated mice; Dox+: doxycycline-treated mice without showing lung tumor; Cancer, doxycycline-treated mice with lung tumor. Mean ± SD in BALF (n>13), DOX -: 8.8±10.0, DOX +: 100.0±49.0, Cancer: 151.8±67.3. Mean ± SD in serum (n>13), DOX -: 6.2±3.7, DOX +: 33.5±35.6, Cancer: 159.0±90.0. The gray lines represent the mean values.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques:

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: Left: CHI3L1 protein concentrations in BALF and serum of doxycycline-treated and untreated bitransgenic mice. Right: ROC (Receiver Operating Characteristic) curve analyses to determine the area under the curve (AUC). Dox-: doxycycline-untreated mice; Dox+: doxycycline-treated mice without showing lung tumor; Cancer, doxycycline-treated mice with lung tumor. A) CCSP-rtTA/(TetO) 7 -CMV-MMP12 mice. Mean ± SD in BALF (n>7), DOX -: 49.6±22.8, DOX +: 49.4±18.1, Cancer: 159.7±22.4. Mean ± SD in serum (n>7), DOX -: 18.6±10.9, DOX +: 18.2±4.6, Cancer: 61.4±33.5. The gray lines represent the mean values; B) CCSP-rtTA/(TetO) 7 -CMV-Api6 mice. Mean ± SD in BALF (n>3), DOX -: 82.4±37.1, DOX +: 93.8±23.0, Cancer: 409.8±49.2. Mean ± SD in serum (n>3), DOX -: 14.0±7.2, DOX +: 30.7±43.1, Cancer: 295.2±194.0. The gray lines represent the mean values.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques:

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: Left: CHI3L1 protein concentrations in BALF and serum of doxycycline-treated and untreated bitransgenic mice. Right: ROC (Receiver Operating Characteristic) curve analyses to determine the area under the curve (AUC). Dox-: doxycycline-untreated mice; Dox+: doxycycline-treated mice without showing lung tumor; Cancer, doxycycline-treated mice with lung tumor. A) c-fms-rtTA/(TetO) 7 -CMV-MMP12 mice. Mean ± SD in BALF (n>7), DOX -: 17.7±13.8, DOX +: 42.1±67.9, Cancer: 70.9±36.5. Mean ± SD in serum (n>7), DOX -: 39.1±25.6, DOX +: 57.7±55.5, Cancer: 232.3±110.6. The gray lines represent the mean values; B) c-fms-rtTA/(TetO) 7 -CMV-Api6 mice. Mean ± SD in BALF (n>6), DOX -: 69.6±16.6, DOX +: 73.2±28.1, Cancer: 189.9±47.2. Mean ± SD in serum (n>6), DOX -: 25.2±7.3, DOX +: 21.0±11.7, Cancer: 69.5±19.1. The gray lines represent the mean values.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques:

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: CCSP-rtTA/(TetO) 7 -CMV-Stat3C bitransgenic mice were treated (+DOX) or untreated (-DOX) with doxycycline. Immunofluorescent staining of lung sections was performed using anti-CHI3L1 and F4/80 antibodies. Co-localization of both staining was observed in the merged picture. Bar represents 20 µm.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques: Staining

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: A) Purity of recombinant CHI3L1-Flag fusion protein (upper arrow); B) LLC cell proliferation treated with GST or CHI3L1-GST fusion protein; C) The apoptotic activity of LLC cells treated with GST or CHI3L1-GST fusion protein by Annexin V labeling assay. *, p<0.05, **, P<0.01.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques: Recombinant, Activity Assay, Labeling

Journal: PLoS ONE

Article Title: Stat3 Downstream Gene Product Chitinase 3-Like 1 Is a Potential Biomarker of Inflammation-induced Lung Cancer in Multiple Mouse Lung Tumor Models and Humans

doi: 10.1371/journal.pone.0061984

Figure Lengend Snippet: Top: CHI3L1 protein concentrations in human serum. Bottom: ROC (Receiver Operating Characteristic) curve analysis to determine the area under the curve (AUC). Mean ± SD in control (n>30): 17.3±7.8. Mean ± SD in adenocarcinoma (n>30): 66.9±64.8. Mean ± SD in squamous carcinoma (n>30): 69.3±69.0. Mean ± SD in small cell cancer (n>30): 56.3±58.8. The gray lines represent the mean values. Control: Normal human without cancer.

Article Snippet: For CHI3L1 protein detection, BALFs from doxycycline-treated or untreated CCSP-rtTA/(TetO) 7 -CMV-Stat3 bitransgenic mice were collected as described above and diluted with Laemmli sample buffer (Bio-Rad, Hercules, CA) at 1∶1, then heated in boiling water for 5 min. Gel electrophoresis and antibody hybridization were performed following the same procedure as above except that rat anti-mouse ChI3L1 primary antibody (1∶1,000, R&D, Minneapolis, MN) and goat anti rat IgG secondary antibody (1∶3000, Vector Laboratories, Burlingame, CA) were used.

Techniques:

Journal:

Article Title: Role for the Related Poly(ADP-Ribose) Polymerases Tankyrase 1 and 2 at Human Telomeres

doi: 10.1128/MCB.22.1.332-342.2002

Figure Lengend Snippet: Analysis of stable cell lines expressing FN-tankyrase 1.WT or HE/A. (A and B) Southern blot analysis of HinfI/RsaI-digested genomic DNA from telomerase-positive HTC75 cell lines (A) or telomerase-negative WI38 cell lines (B) expressing vector control (V), FN-tankyrase 1.WT (WT), or FN-tankyrase 1.HE/A (HE/A). Cell lines were grown for 88 (HTC75) or 19 (WI38) population doublings (PD), and DNA samples were analyzed at the indicated PDs. Blots were probed with TTAGGG-repeat probe to detect telomeric restriction fragments. (C and D) Graphical representations of telomere length changes in HTC75 (C) or WI38 (D) cell lines expressing FN-tankyrase 1.WT, HE/A, or vector. Plots represent the mean telomere length values derived from the Southern blots analyzed in panels A and B. (E) Western blot analysis of whole-cell extracts from HTC75 (PD 52) or WI38 (PD 4) cells expressing vector control (V), FN-tankyrase 1.WT (WT), or FN-tankyrase 1.HE/A (HE/A). Blots were probed with the following antibodies: anti-tankyrase 1 376, anti-poly(ADP-ribose), anti-TRF1 415, and anti-TRF2.

Article Snippet: HTC75 cells (an HT1080-derived clonal cell line [ 40 ]) or WI38 cells (human primary fibroblasts at population doubling [PD] 30; American Type Culture Collection) were infected essentially as described ( 32 ).

Techniques: Stable Transfection, Expressing, Southern Blot, Plasmid Preparation, Derivative Assay, Western Blot