Journal: The FASEB Journal

Article Title: Suppression of cigarette smoke induced MMP1 expression by selective serotonin re‐uptake inhibitors

doi: 10.1096/fj.202001966RR

Figure Lengend Snippet: Validation of and NMLSMR targets identified by the high throughput assay. A, To test for reproducibility of the assay, 293T cells were transfected with the MMP‐1/pGL3 reporter plasmid. Cells were seeded in three 96‐well plates using an interleaved format and treated with 5% CSE media (high‐signal) 2% CSE (medium‐signal) no CSE (low‐signal). After a 24‐h incubation, the luciferase activity was measured in each well. Representative data from days 1–3 is shown. The calculated Z' factor was 0.71 ± 0.05 SEM, n = 5. B, As expected, the ERK inhibitor PD98059 inhibited luciferase expression in a dose‐dependent fashion in positive controls exposed to 5% CSE ( *** p ≤ 0.001, significantly different from control; ### p ≤ 0.001, significantly different from promoter + CSE; ## p ≤0.01, significantly different from promoter + CSE). C, Data from the high throughout screen displaying each compound tested as a block dot. The y‐axis illustrates the change in MMP1 expression compared with expression levels in CSE exposed in HEK293 cells. The drugs in the −100% range (red shaded area) are capable of nullifying all CSE induced MMP1 transcriptional activity

Article Snippet: MMP‐1 Fluorokine E kits (R&D Systems F1M00) were used to determine MMP‐1 levels and performed per manufacturer's protocol.

Techniques: Biomarker Discovery, High Throughput Screening Assay, Transfection, Plasmid Preparation, Incubation, Luciferase, Activity Assay, Expressing, Control, Blocking Assay

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 1: Role in Sarcoma Biology

doi: 10.1371/journal.pone.0014250

Figure Lengend Snippet: Figure 2A) Relative quantity (RQ) of MMP1 in the stable clones of 143B cell-line prepared. MMP1 Silenced is 98% down-regulated as compared to control. Control = 1.00±0.13; MMP1 Silenced = 0.18±0.14. Figure 2B) displays the Western Blot analysis for MMP1 production by the stable clones of 143B cell-line. MMP1 Silenced clone shows down-regulation at the level of protein as compared to control. Figure 2C) displays in vitro proliferation assay showing growth of control and MMP1 silenced clones over 5 days. There is an approximately 14% increased growth displayed by MMP1 silenced clone in the log-phase of growth. Difference in mean growth over 5 days is statistically insignificant (p = 0.804). Figure 2D) displays results for the in vitro invasion assay measured in Relative Light Units (RLU). Control = 21272.7±790; MMP1 silenced = 8008.1±874.5 RLUs (p = 1.54E-09).

Article Snippet: The stable clones were generated by pHuSH 29mer shRNA constructs against MMP1 (#TR311450 RNAi targeting vector system OriGene Rockville, MD) as described .

Techniques: Clone Assay, Western Blot, In Vitro, Proliferation Assay, Invasion Assay

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 1: Role in Sarcoma Biology

doi: 10.1371/journal.pone.0014250

Figure Lengend Snippet: Tumor volumes in ‘control’ and ‘MMP1 silenced’ groups at 5 week intervals are shown in Figure 3A. Figure 3B displays the histograms showing mean volume of tumor (mm 3 ) for a single mouse in control and MMP1 silenced groups at corresponding time points. Difference in tumor volumes at 2, 4 and 5 weeks is significant with p-values of 0.023, 0.036 and 0.026.

Article Snippet: The stable clones were generated by pHuSH 29mer shRNA constructs against MMP1 (#TR311450 RNAi targeting vector system OriGene Rockville, MD) as described .

Techniques:

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 1: Role in Sarcoma Biology

doi: 10.1371/journal.pone.0014250

Figure Lengend Snippet: Confocal Images showing primary tumor from a) control group and b) MMP1 Silenced group. Images c) and d) display only the blood vessels from a) and b). Figure 4E shows histograms representing average vascular volume (number of red pixels) for a single field of vision (at 25×) in tumors harbored by control (183829.7±78022) and MMP1 Silenced (323147.5±41141) groups. Difference in mean vascular volume is significant with p-value = 0.01.

Article Snippet: The stable clones were generated by pHuSH 29mer shRNA constructs against MMP1 (#TR311450 RNAi targeting vector system OriGene Rockville, MD) as described .

Techniques:

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 1: Role in Sarcoma Biology

doi: 10.1371/journal.pone.0014250

Figure Lengend Snippet: Lungs showing metastatic areas from control (Figure 5A) and MMP1 silenced groups (Figure 5B) are shown. Figure 5C shows histogram representing average percentage of total lung volume affected by metastasis at 5 weeks for control (54±17.6%) and MMP1 Silenced (36±23.7%) groups. Difference in mean percentage area affected by metastasis is not significantly different, p = 0.08.

Article Snippet: The stable clones were generated by pHuSH 29mer shRNA constructs against MMP1 (#TR311450 RNAi targeting vector system OriGene Rockville, MD) as described .

Techniques:

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 1: Role in Sarcoma Biology

doi: 10.1371/journal.pone.0014250

Figure Lengend Snippet: Figure 6A shows the Relative quantity (RQ) of MMP1 in the control and MMP1 silenced clones isolated from mouse lungs at necropsy. MMP1 Silenced clone is 95% down-regulated as compared to control. Control = 1.72±0.17; MMP1 Silenced = 0.05±0.08. Figure 6B shows MMP1a production by peri-tumoral murine stromal tissue as assessed with laser capture microdissection and QPCR.

Article Snippet: The stable clones were generated by pHuSH 29mer shRNA constructs against MMP1 (#TR311450 RNAi targeting vector system OriGene Rockville, MD) as described .

Techniques: Clone Assay, Isolation, Laser Capture Microdissection

Journal: Cancers

Article Title: CD83, a Novel MAPK Signaling Pathway Interactor, Determines Ovarian Cancer Cell Fate

doi: 10.3390/cancers12082269

Figure Lengend Snippet: Integrated transcriptome and proteome analyses. ( a ) Volcano plots show the differentially expressed genes of NC-vs.-KD and NC-vs.-OV, as revealed by transcriptome. ( b ) CD83-KD SKOV3 cells exhibited elevated expression of matrix metallopeptidases (MMPs) and TGF-beta family members as compared with NC and CD83-OV cells. ( c ) The expression of cell cycle regulators and stemness-related genes was upregulated in CD83-OV SKOV3 cells. ( d ) The intersection of the transcriptome (fold change ≥2) and proteome (fold change ≥1.5) identified 83 DE proteins (and protein-coding genes) between CD83-KD and NC SKOV3 cells. Correlation analysis of DE genes and DE proteins indicated the consistency (R = 0.5360) between transcriptome and proteome in this study. ( e ) Venn diagram shows the downregulated proteins (e.g., CD83, KIT) and upregulated proteins (e.g., MMP1, MMP7) in the CD83-KD SKOV3 cells. ( f ) Content of MMP1 and MMP7 in SKOV3 cell lysate and culture media by ELISA assay. The differences between KD and NC, as well as KD and OV, were presented as the mean±SEM; Student’s t -test; ** p < 0.01.

Article Snippet: MMP1 and MMP7 concentrations from cell lysate and cell culture supernatant were measured using MMP1 ELISA Kit (Abcam, Cambridge, MA, USA) and MMP7 ELISA Kit (CUSABIO, Wuhan, China), respectively, following the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay