Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 1 Harnessing fibroblast contractility to engineer dense carcinoma constructs. (a) Schematic illustration of fibroblast‐driven contraction of collagen fibers around cancer spheroids. Blue shapes represent fibroblasts, black lines represent collagen fibers, and green clusters represent spheroids. (b) Side by side comparison of H&E‐stained sections of dense breast carcinoma constructs (left) and biopsied mammary ductal carcinoma tissue (right). Scale bars = 80 μm. (c) Time‐course imaging of gross contraction and compaction of dense carcinoma constructs. Scale bars = 200 μm. (d) Box plot of the relative area change over time for gels seeded at 2.5 × 105 HLF/mL (n = 6) and 5 × 105/mL (n = 7). **p < 0.01. ***p < 0.001. See Section 2 for details of statistical analyses. (e) Box plot of the corresponding increase in estimated collagen density over time for gels seeded for acellular gels and 5 × 105 HLF/mL (n = 5 per group).

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Construct, Comparison, Staining, Imaging

Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 2 Effect of fibroblast cell type and seeding density on tissue contraction. (a–c) Time‐course imaging of gross contraction and compaction of dense carcinoma constructs formed using several types of fibroblasts. Constructs were seeded at 5 × 105/mL (n = 3 per group). Scale bars = 3 mm. Similar kinetics and magnitude of contraction using human dermal fibroblasts (HDF, a), human lung cancer associated fibroblasts (CAF, b), and human lung fibroblasts (HLF, c). (d and e) Time‐course imaging of gross contraction and compaction for up to 3 weeks in culture. Scale bars = 3 mm. Constructs were seeded at 0.5 × 105 HLF/mL (n = 6, panel d) and 5 × 105/mL (n = 7, e). (f) Relative area change over time as a measure of contraction for each type of fibroblast during 7 days of culture as shown in (a–c). No significant differences were measured. HDF constructs were slightly less contracted at 7 days, matching the qualitatively larger size in time course images. (g) Relative area change over time as a measure of contraction. Similar final volumes were reached with each seeding density. Contraction was more gradual with a seeding density of 0.5 × 105 HLF/mL.

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Imaging, Construct

Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 5 Local anisotropy is similar in constructs formed with various types of contractile cells. (a) H&E‐staining of sectioned dense carcinoma constructs formed with cancer associated fibroblasts (CAF), human dermal fibroblasts (HDF), and human lung fibroblasts (HLF). Seven days, scale bars = 30 μm. (b) Collagen type I staining of sectioned dense carcinoma constructs formed with CAF, HDF, and HLF. Seven days, scale bars = 20 μm. (c) Box plot of the relative fluorescent intensity in binned regions of interest at the specified distances from spheroid for dense carcinoma constructs formed with CAF (white), HDF (light gray), and HLF (dark gray) (n = 9 per group). *p < 0.05, **p < 0.01. One way ANOVA. (d) Box plot of fiber orientation relative to spheroid tangent line and distance from spheroid for dense carcinoma constructs. Same shading as (d) to indicate fibroblast type; CAF (white), HDF (light gray), and HLF (dark gray) (n = 9 per group). One way ANOVA. ns, not significant.

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Construct, Staining

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) A schematic outlining the strategy for SILAC-based identification of SASP proteins. See for time lines of how cells were made quiescent or senescent and how quiescent and senescent conditioned media (CM) were produced. (B) Reactome pathway analysis of proteins secreted at >2-fold levels by senescent compared with quiescent HCA2 cells. Pathways related to hemostasis are identified by boxes. (C) Heatmap showing individual levels of proteins, identified by SILACs, that participate in hemostasis and are secreted by quiescent (QUI) and senescent (SEN [IR]) HCA2 normal human fibroblasts. (D) CM were collected from quiescent (QUI) and senescent (SEN [IR]) cells, concentrated, and analyzed using western blotting for the indicated SASP factors. Cells remaining on the plate at the time of CM collection were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. Fold changes are mean ± SEM. *p < 0.05 and **p < 0.01. See also and .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Multiplex sample analysis, Produced, Western Blot, Staining

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) mRNA was extracted from HCA2 cells made quiescent (QUI) or induced to senesce by doxorubicin (SEN [DOXO]), and analyzed using qPCR for the indicated genes, using actin mRNA for normalization. The data are shown in a.u., and the bars show mean ± SEM. *p < 0.05 and **p < 0.01, two-tailed, unpaired t test. (B) CM were collected from cells treated as in (A), concentrated and analyzed using western blotting for the indicated SASP factors. Cells were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. (C) IMR-90 fibroblasts were depleted of mtDNA by culture in ethidium bromide until senescent (21 days) (labeled as MiDAS). RNA was extracted and analyzed using qPCR for the indicated factors. Heatmap represents individual values for three experiments. *p < 0.05, one-way ANOVA. See also .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Two Tailed Test, Western Blot, Staining, Labeling

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet:

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Recombinant, Modification, Sequencing, Multiplex sample analysis, Labeling, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Reverse Transcription, Software

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 5. RAGE immunoblotting analysis of human intestine. SDS-PAGE of lung lysate (1.5 μg/lane) as well as lysates from small intestine (SI) and colon (50 μg/lane) was performed under reducing conditions. Membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 4. (A) RAGE immunoblotting analysis of human lung lysate. SDS-PAGE of lung lysate (1 μg/lane; PNGase F -) and deglycosylated lung lysate (1 μg/lane; PNGase F +) was performed under reducing conditions. Membranes were stained with four different anti-RAGE antibodies. Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. (B) RAGE immunoblotting analysis of RAGE overexpressing (+) and control (−) HEK293T cells. SDS-PAGE was performed under reducing conditions. For WB analysis membranes were stained with anti-FLAG and anti-RAGE (AB D01P) antibodies. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 6. RAGE immunoblotting analysis of human placenta derived from different healthy (H) as well as GDM-, FGR- and PE-affected pregnancies. SDS-PAGE of lung lysate (1 μg/lane) as well as lysates from the placentas (25 μg/lane) was performed under reducing conditions. For WB analysis membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). Ponceau S staining was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, Derivative Assay, SDS Page, Staining, Negative Control, Control

Journal: Oncotarget

Article Title: MicroRNA-224 is implicated in lung cancer pathogenesis through targeting caspase-3 and caspase-7.

doi: 10.18632/oncotarget.5224

Figure Lengend Snippet: Figure 2: The expressions of CASP3 and CASP7 mRNA are down regulated in NSCLC. a. CASP7 mRNA expressions in thirty paired NSCLC tissues and NATs. b. CASP3 mRNA expressions in thirty paired NSCLC tissues and NATs. GAPAH was used for normalization. c. and d. Kaplan-Meier plots of overall survival of lung cancer patients, stratified by expression of CASP7 (1926 patients) c. or CASP3 (1926 patients) d.. Data obtained from the Kaplan-Meier plotter database (kmplot.com/analysis).

Article Snippet: nsclc tissue samples Human Lung Tissue Microarrays (IMH-358) were purchased from Novus Biologicals, San Diego, CA.

Techniques: Expressing

Journal: Oncotarget

Article Title: MicroRNA-224 is implicated in lung cancer pathogenesis through targeting caspase-3 and caspase-7.

doi: 10.18632/oncotarget.5224

Figure Lengend Snippet: Figure 4: Expressions of CASP3 and CASP7 are inversely correlated with miR-224. a. Representative pictures and summary of co-expression analyses for miR-224 and CASP7 in NSCLC tissues. MiR-224 was detected by using 5’-DIG labeled LNA probe (purple) and CASP7 was detected by immunohistochemistry (red). Left: High CASP7 and low/neg miR-224, Middle: similar expression of CASP7 and miR-224, right: Low/neg CASP7 and high miR-224. b. Representative pictures and summary of co-expression analyses for miR- 224 and CASP3 in NSCLC tissues. MiR-224 was detected by using 5’-DIG labeled LNA probe (purple) and CASP3 was detected by immunohistochemistry (red). Left: High CASP3 and low/neg miR-224, Middle: similar expression of CASP3 and miR-224, right: Low/ neg CASP3 and high miR-224. c. In the cases of NSCLC where both miR-224 and CASP7 expression were noted, no detectable CASP7 was found in cancer cells overexpressing miR-224 (purple arrow).

Article Snippet: nsclc tissue samples Human Lung Tissue Microarrays (IMH-358) were purchased from Novus Biologicals, San Diego, CA.

Techniques: Expressing, Labeling, Immunohistochemistry

Journal: bioRxiv

Article Title: Sperm-specific COX6B2 enhances oxidative phosphorylation, proliferation, and survival in lung adenocarcinoma

doi: 10.1101/2020.04.09.030403

Figure Lengend Snippet: (A-C) Kaplan-Meier curves for OS and FP in NSCLC (A) LUAD (B) and LUSC patients (C). (D) COX6B2 mRNA expression (RNA-seq RSEM, log2(norm count +1)) from TCGA Lung Cancer dataset. Bars represent median (normal: n = 110; tumor: n = 1017). P values calculated by Mann-Whitney test. (E-F) Whole tissue homogenates of LUAD tumors (E) and LUAD cell line lysates (F) were immunoblotted with indicated antibodies. Molecular weight (MW) markers are indicated. (G) IHC staining of non-malignant testis (a positive control), non-malignant lung (adjacent normal) and LUAD tissues. Scores ranged from 0 to 3. Scale bar, 50 μm. Bars represent mean ± SEM. P values calculated by Mann-Whitney test. (H) Representative confocal images of endogenous COX6B2 in HCC515 cells. Tom20 is used as a mitochondrial marker. Images were shown as Z-stack maximum projection from 0.3 μm-thick-image. Scale bar, 10 μm.

Article Snippet: Human normal lung whole tissue lysate was purchased from Novus Biologicals (NB820-59237).

Techniques: Expressing, RNA Sequencing, MANN-WHITNEY, Molecular Weight, Immunohistochemistry, Positive Control, Marker

Journal:

Article Title: Phosphorylated Eukaryotic Translation Initiation Factor 4 (eIF4E) is Elevated in Human Cancer Tissues

doi:

Figure Lengend Snippet: Comparison of p-eIF4E expression between the tumors and adjacent normal tissues

Article Snippet: Human lung cancer TMA consisting of 40 cases of stage I-III lung cancer tissues, 10 cases of metastatic cancer tissues from the primary lung cancer, and 9 cases of adjacent normal human lung tissues was purchased from Imgenex (IMH-358; SanDiego, CA).

Techniques: Comparison, Expressing, Wilms Tumor Assay

Journal: Translational Oncology

Article Title: Vasorin/ATIA Promotes Cigarette Smoke–Induced Transformation of Human Bronchial Epithelial Cells by Suppressing Autophagy-Mediated Apoptosis

doi: 10.1016/j.tranon.2019.09.001

Figure Lengend Snippet: Increased vasorin expression in human lung cancer and HBECs exposed to tobacco carcinogens . (A) Tissue arrays were stained for vasorin with immunohistochemistry. The normal airway and alveolar epithelial cells are weakly positive while the tumor cells are strongly positive. (B) Results of tissue arrays with comparison of the staining between vasorin and normal lung epithelial cells. (C) Vasorin was examined in nontransformed (HBEC-1 and -2) and lung cancer cell lines. Relative vasorin fold increase (HBEC-2 was set to 1) was shown. (D) BEAS-2B and HBEC-2 cells were treated with CSE for 24 hours. (E) HBEC-2B (BPDE-transformed) cells were compared with HBEC-2 cells for vasorin expression. Protein was detected by western blot in C to E with β-actin as loading control.

Article Snippet: Immunohistochemistry staining using VECTASTAIN ABC Kit and DAB (3,3′-diaminobenzidine) Peroxidase Substrate Kit (Vector Laboratories, Burlingame, CA) and result assessment of human lung cancer tissue array (Imgenex; Novus Biologicals, Centennial, CO) has been described previously [ ].

Techniques: Expressing, Staining, Immunohistochemistry, Comparison, Transformation Assay, Western Blot, Control