Journal:

Article Title: Collagen XXIII--a potential biomarker for the detection of primary and recurrent non-small cell lung cancer

doi: 10.1158/1055-9965.EPI-09-1095

Figure Lengend Snippet: Collagen XXIII immunohistochemical analysis of multiple cancer tissues

Article Snippet: Antibody Validation A new human collagen XXIII mouse monoclonal antibody was made against the cleaved portion of human collagen XXIII by R&D Systems.

Techniques: Immunohistochemical staining, Staining

Journal:

Article Title: Collagen XXIII--a potential biomarker for the detection of primary and recurrent non-small cell lung cancer

doi: 10.1158/1055-9965.EPI-09-1095

Figure Lengend Snippet: Representative images showing collagen XXIII staining in the NSCLC subtypes: A) lung adenocarcinoma, B) large cell carcinoma, and C) squamous cell carcinoma. D) Collagen XXIII staining is absent in small cell lung cancer tissues. Staining was performed using the 468642 antibody and images were acquired using a 20X objective lens.

Article Snippet: Antibody Validation A new human collagen XXIII mouse monoclonal antibody was made against the cleaved portion of human collagen XXIII by R&D Systems.

Techniques: Staining

Journal:

Article Title: Collagen XXIII--a potential biomarker for the detection of primary and recurrent non-small cell lung cancer

doi: 10.1158/1055-9965.EPI-09-1095

Figure Lengend Snippet: A) Bar graph showing percentage of patient tumor or normal tissue cores expressing collagen XXIII across all NSCLC TMAs and each individual TMA. Single asterisks indicate unavailable data. Patients lacking detectable collagen XXIII staining in all of their representative tissue cores were considered negative for collagen XXIII protein expression (Supplementary Table 1). Chi-squared analysis of a 3 by 2 contingency table for the combined TMA data was highly significant (p<0.001). A two-tailed p-value was calculated for each comparison between NSCLC subtype and normal lung tissue using Fisher’s Exact Test. Highly significant differences (p<0.001) are indicated (***). For immunohistochemical analysis, the 468642 antibody was used on the US Biomax and M.D. Anderson Cancer Center TMAs, while the 6.1B antibody was used on the Brigham and Women’s Hospital TMA. B) ROC curve plotting the true positive rate against the false positive rate for the different possible collagen XXIII staining intensity thresholds. (Tabulated data in supplementary table 2) Area under the curve = 0.860. C) Representative Kaplan Meier curves demonstrating reduced recurrence-free survival in patients with high (>85) collagen XXIII staining intensity (p= 0.013). Data for all examined threshold values is included in supplementary table 4.

Article Snippet: Antibody Validation A new human collagen XXIII mouse monoclonal antibody was made against the cleaved portion of human collagen XXIII by R&D Systems.

Techniques: Expressing, Staining, Two Tailed Test, Comparison, Immunohistochemical staining

Journal:

Article Title: Collagen XXIII--a potential biomarker for the detection of primary and recurrent non-small cell lung cancer

doi: 10.1158/1055-9965.EPI-09-1095

Figure Lengend Snippet: A) Collagen XXIII is detected in urine from 23 of 29 (79%) NSCLC patients but only in 15 of 54 (28%) control urine samples (Fisher’s Exact test; p<0.0001 ***). B) Representative Western blot demonstrating collagen XXIII positivity in a subset of urine samples using the 468642 antibody. Recombinant collagen XXIII corresponding to the cleaved ectodomain was used as a positive control (+ con) and is the predominant form of urinary collagen XXIII. Arrow indicates potential non-reduced protein complexes.

Article Snippet: Antibody Validation A new human collagen XXIII mouse monoclonal antibody was made against the cleaved portion of human collagen XXIII by R&D Systems.

Techniques: Control, Western Blot, Recombinant, Positive Control

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: COL23A1 expression in ccRCC tissues and paired ANTs. ( A ) Results of qRT-PCR exhibited the relative mRNA levels of COL23A1 in primary non-metastatic ccRCC tissues compared with paired ANTs from FUSCC cohort. Error bars are ± SD (n = 3). ( B ) The expression of COL23A1 mRNA in primary non-metastatic ccRCC tissues compared with paired ANTs from TCGA cohort. ( C ) Results of western blot analysis showed the COL23A1 protein levels in primary non-metastatic ccRCC tissues compared with paired ANTs from FUSCC cohort.

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: The correlation between COL23A1 expression and prognosis in ccRCC. ( A , B ) Representative high COL23A1 level in ccRCC tissues with ( A ) ×200 magnification and ( B ) ×400 magnification. ( C,D ) Representative low COL23A1 level in ccRCC tissues with ( C ) ×200 magnification and ( D ) ×400 magnification. ( E,F ) Representative COL23A1 level in normal kidney tissues with ( E ) ×200 magnification and ( F ) ×400 magnification. ( G ) Kaplan-Meier curves for OS after radical nephrectomy.

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: Clinicopathological characteristics in relation to COL23A1 expression status.

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of OS in 151 ccRCC patients.

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: The mRNA and protein expression of COL23A1 in normal and ccRCC cell lines. ( A ) A statistical analytical graph showed COL23A1 mRNA levels in the human kidney cortex cell lines (HKC) and four ccRCC cell lines (786-O, A-498, Caki-1, OS-RC-2). Error bars are ± SD (n = 3). ( B ) Results of western blot analysis showed COL23A1 protein levels in the human kidney cortex cell lines (HKC) and four ccRCC cell lines (786-O, A-498, Caki-1, OS-RC-2).

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: COL23A1 was down-regulated by small interfering RNA (siRNA) in ccRCC cell lines. ( A , B ) Relative COL23A1 mRNA levels in (A) 786-O and ( B ) A-498 cells that were transfected with siRNAs (#1, #2, #3) compared with siNC respectively were analyzed using qRT-qPCR. Error bars are ± SD (n = 3). ( C ) Results of western blot analysis showed the COL23A1 protein level of 786-O cells that were transfected with siRNAs (#1, #2, #3) compared with siNC, and the histogram right was quantification of the western blot analysis results. Error bars are ± SD (n = 3). ( D ) Results of western blot analysis showed the COL23A1 protein level of A-498 cells that were transfected with siRNAs (#1, #2, #3) compared with siNC, and the histogram right was quantification of the western blot analysis results. Error bars are ± SD (n = 3).

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Small Interfering RNA, Transfection, Western Blot

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: Effect of COL23A1 silencing on cell proliferation and cell cycle in ccRCC cells. ( A ) CCK-8 assays showed the effect of COL23A1 knockdown on the proliferation of 786-O cells. Error bars are ± SD (n = 3). ( B ) CCK-8 assays showed the effect of COL23A1 knockdown on the proliferation of A-498 cells. Error bars are ± SD (n = 3). ( C ) Effect of COL23A1 silencing on the cell cycle of 786-O cells. Error bars are ± SD (n = 3). ( D ) Effect of COL23A1 silencing on the cell cycle of A-498 cells. Error bars are ± SD (n = 3).

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: CCK-8 Assay, Knockdown

Journal: Scientific Reports

Article Title: The Oncogenic Role of COL23A1 in Clear Cell Renal Cell Carcinoma

doi: 10.1038/s41598-017-10134-2

Figure Lengend Snippet: Effect of COL23A1 silencing on cell migration and adhesion in ccRCC cells. (A, B) Effect of COL23A1 silencing on the cell adhesion of (A) 786-O and (B) A-498 cells. Error bars are ± SD (n = 3). (C, D) Effect of COL23A1 silencing on the expression of proteins involved in cell adhesion of (C) 786-O and (D) A-498. (E) Representative images of migration assays performed using 786-O (upper left) and A-498 (lower left) cells (scale bars, 100 µm). The histograms right were quantification of the migration assays of 786-O (upper) and A-498 (lower) cells. Error bars are ± SD (n = 3).

Article Snippet: Slides were blocked using normal goat serum followed by the avidin/biotin blocking kit and incubated with anti-COL23A1 antibody (1:200, Catalog # MAB4165, R&D Systems, Minneapolis, Minnesota, USA) at 4 °C overnight.

Techniques: Migration, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Methoxyflavonoid Isosakuranetin Suppresses UV-B-Induced Matrix Metalloproteinase-1 Expression and Collagen Degradation Relevant for Skin Photoaging

doi: 10.3390/ijms17091449

Figure Lengend Snippet: Effect of isosakuranetin on the level of type-1 procollagen in UV-B-irradiated NHDF cells. After pretreatment of fibroblast cells with different concentrations of isosakuranetin for 24 h, cells were irradiated with UV-B (20 mJ/cm 2 ), and then cells were further incubated for 24 h. Total collagen levels were measured in the culture media of control cells or UV-B-treated cells by SirCol assay. Data shown are mean values ± S.D. * p < 0.05 vs. non-UV-B-irradiated cells, ** p < 0.05 vs. UV-B-irradiated cells.

Article Snippet: Antibodies against human MMP-1 and human procollagen were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Irradiation, Incubation, Control