specific mmp2 inhibitor  (Merck & Co)

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 1. Treatment with the MMP2/9 inhibitor SB3CT during the MD period suppressed OD plasticity in adult mouse V1. A, Experimental timeline testing the effect of selective MMP2 and MMP9 inhibition on visual plasticity in healthy adult mice. Between P70 and P110, a group of mice received MD. From d0 (before MD), mice were checked daily for 7 d in the optomotor setup for the assessment of both spatial frequency and contrast sensitivity thresholds. One hour after MD, mice of all experimental groups were injected intraperitoneally with SB3CT or vehicle solu- tion; injections continued until d7, when V1 activity was visualized using intrinsic signal optical imaging. B–E, Representative activity maps of the left binocular V1 and ODI values of vehicle- injected (B, D) or SB3CT-injected (C, E) mice. Greyscale-coded response magnitude maps, polar maps, two-dimensional OD maps, and histograms of OD scores including the average ODIs are illustrated. In mice without MD, activity patches evoked by stimulation of the contralateral eye were darker than those of the ipsilateral eye, the average ODI was positive, and warm colors pre- vailed in the OD maps, indicating contralateral dominance (B, C). In vehicle-injected mice, 7-d MD induced a strong OD shift toward the nondeprived (ipsilateral) eye (D): ipsilateral eye stimula- tion now caused darker V1-activity patches compared with noMD mice, colder colors appeared in the OD map, and average OD scores shifted to the left. In contrast, SB3CT-injected mice did not display OD plasticity (E). After MD, the contralateral eye continued to dominate V1 activation, warm colors continued to prevail in the OD map, and the average ODI was not changed (red) in SB3CT-treated mice (E). Scale bars: 1 mm.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Inhibition, Injection, Activity Assay, Optical Imaging, Activation Assay

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 2. Quantification of ODIs (A) and V1 activation (B) in vehicle-injected (black) and SB3CT-injected (green) mice with or without MD. A, Administration of SB3CT for 7 d abol- ished OD plasticity in MD mice. Symbols represent ODI values of individuals; filled circles rep- resent mice without MD and half-filled circles mice with MD; mean values are shown as horizontal lines. B, V1 activation (fractional change in reflectance 104) elicited by stimu- lation of the contralateral (C) or ipsilateral (I) eye without and after MD (deprived eye is marked by a black filled circle on the x-axis). In vehicle-treated mice, the OD shift was pri- marily mediated by an increase of open (ipsi) eye responses in V1, while after SB3CT treat- ments, there was no significant increase in open eye responses of MD mice receiving injections. Multivariate ANOVA, followed up by Sidak-corrected multiple comparisons. Mean 6 SEM, *p , 0.05, **p , 0.01, ***p , 0.001. Tables comparing the two independent analyses are provided as the Extended Data Figure 2-1.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Activation Assay, Injection

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 3. Daily injection of the MMP2/9-specific inhibitor SB3CT increased PNNs in V1, stained with WFA. A, WFA-positive staining of PNNs in vehicle-injected (left) and SB3CT- injected (right) mice. PNNs were identified by their shape and bright circular outline through- out the layers of V1. One example of each condition is displayed. On the right, noMD/MD conditions of vehicle-injected mice are shown. On the left, noMD/MD conditions of SB3CT- inejcted mice are shown. Scale bar: 100 mm. B, Quantification of PNNs in V1. Number of WFA-positive PNNs were significantly higher in SB3CT-injected mice, regardless of MD. From each brain, three sections containing V1 were analyzed. ANOVA followed up by Sidak-cor- rected test for simple effects. Scale bar: 100 mm. Mean 6 SEM, *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Injection, Staining

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 4. Injection of the MMP2/9 inhibitor SB3CT prevented experience-induced enhancements of both the spatial frequency (A) and contrast sensitivity (B) thresholds of the optomotor reflex of the open eye in adult mice after MD (half-filled circles). A, Spatial frequency threshold values in cyc/° are plotted against days, for mice without (filled circles) and with MD (half-filled circles) for both vehicle-treated (black) and SB3CT-treated (green) mice. After 7 d of MD, spatial frequency thresholds improved significantly in vehicle-injected MD mice when compared with mice without MD and SB3CT-injected MD mice. In contrast, SB3CT-injected MD mice did not significantly increase thresholds until D7, while control MD mice improved significantly each day. Mice without MD did not show any change over days. Three-way mixed ANOVA was used, followed up by Sidak-corrected tests for simple effects. B, In vehicle-treated MD mice, contrast sensitiv- ity thresholds of the open eye increased significantly after 7-d MD compared with both noMD mice (filled circles) and SB3CT-injected noMD mice (blue). SB3CT MD mice (blue, half-filled circles) had significantly lower contrast sensitivity thresholds than vehicle-injected mice (black, half-filled circles), similar to noMD mice (filled circles). Gratings left of the y-axis illustrate the contrast steps for a given (constant) spatial frequency. Four-way mixed ANOVA, followed up by Sidak-corrected tests for simple effects. Mean 6 SEM, *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Injection, Control

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 5. Brief treatment with the MMP2 and MMP9-specific inhibitor SB3CT rescued adult OD plasticity in V1 after a stroke in S1. A, The experimental design to test the effect of MMP2 and MMP9 inhibition on OD plasticity after a PT stroke in S1. A small (1 mm in diameter) lesion in S1 was induced on young adult mice (P70–P90) using PT. Shortly after (1 h), half of the PT mice received MD. One hour after MD, mice were injected with SB3CT or vehicle solution and returned to their home cages. Starting from d0 before MD, mice were checked daily in the optomotor setup for the assessment of spatial frequency and contrast sensitivity thresholds of the optomotor reflex for the following 7d. On d7, optical imaging of intrinsic signals was performed, after which the mice were perfused, and the brains were harvested. After tissue preparation and immunohistochemistry, PT lesions were measured. Representative activity maps of binocular V1 and ODI values of the left hemisphere measured by optical imaging of intrinsic signals in PT-lesioned, one-time vehicle-injected (B, D) or SB3CT-injected (C, E), or sham-treated vehicle-injected (F, H) or SB3CT- injected (G, I), mice are displayed. In PT mice without MD (B, C), activity patches evoked by stimulation of the contralateral eye were darker than those of the ipsilateral eye, the average ODI was positive, and warm colors prevailed in the OD maps, indicating contralateral dominance in the left hemisphere binocular V1. Vehicle-injected PT mice with MD (deprived eye is marked by black filled

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Inhibition, Injection, Optical Imaging, Immunohistochemistry, Activity Assay

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 6. Quantification of OD plasticity on the brief (1-d) inhibition of MMP2/MMP9 with SB3CT after a PT lesion in S1: both ODIs (A) and V1 activation (B) without and with MD in the var- ious treatment groups (vehicle/SB3CT and PT/sham lesion) are illustrated. A, Optically imaged ODIs of mice injected with vehicle (black/gray) or SB3CT (dark green, light green). Symbols repre- sent ODI values of individuals (squares specify PT-lesioned mice, circles sham-lesioned mice). In PT mice with MD, a single systemic administration of SB3CT on d0 rescued the impaired OD shift (impaired by PT in vehicle-treated mice). B, V1 activation elicited by stimulation of the contralateral (C) or ipsilateral (I) eye without and after MD. Multivariate ANOVA, followed up by Sidak-corrected tests for simple effects. Data display as in Figure 2. Mean 6 SEM, *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Inhibition, Activation Assay, Injection

Journal: The Journal of Neuroscience

Article Title: MMP2 and MMP9 Activity Is Crucial for Adult Visual Cortex Plasticity in Healthy and Stroke-Affected Mice

doi: 10.1523/jneurosci.0902-21.2021

Figure Lengend Snippet: Figure 7. Injection of the MMP2/9 inhibitor SB3CT after the induction of a PT lesion in S1 rescued experience-enabled enhancements of both the spatial frequency (A) and contrast sensitivity (B) thresholds of the optomotor reflex in adult mice. A, Spatial frequency threshold values in cyc/° are plotted against days for PT (squares), sham (circles), in mice without (filled symbols) and with MD (half-filled symbols), both for vehicle (black/gray) and SB3CT (dark green, light green) treat- ment. Sevendays of MD induced a significant increase in spatial frequency threshold in SB3CT-injected PT-treated mice each day but not in vehicle-injected PT mice. In contrast, sham-lesioned mice showed significant threshold increases over days after MD (half-filled squares) compared with noMD mice, regardless of the type of injection (vehicle/SB3CT). B, Likewise, SB3CT treatment after PT rescued the experience-induced increase in contrast sensitivity thresholds of the open eye after MD: PT mice receiving a single SB3CT injection (dark green) had higher contrast thresholds, while the contrast thresholds of vehicle-treated PT mice (black) were significantly lower compared with SB3CT-injected PT mice. Sham- lesioned MD mice also had significantly higher contrast thresholds at every spatial frequency compared with noMD mice, regardless of treatment (vehicle and SB3CT). Gratings left of the y-axis illustrate the contrast steps for a given (constant) spatial frequency. Mixed ANOVA followed up by Sidak-corrected test for simple effects. Mean 6 SEM, *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Administration of the gelatinase (MMP2 and MMP9)-specific inhibitor SB3CT To specifically inhibit the activity of MMPs 2 and 9 in the visual cortex, the specific inhibitor SB3CT [chemical name: 2-[[(4-phenoxyphenyl) sulfonyl] methyl]-thiirane; formula: C15H14O3S2; SelleckChem] was used and prepared as previously described with minor modifications as specified below (Brown et al., 2000; Gu et al., 2005).

Techniques: Injection

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells were treated with GA (200 mg/L) or BSA (200 mg/L) for 24 h. Antioxidant NAC was dissolved in sterile double-distilled water and used in different final concentrations with GA (200 mg/L). The antioxidant effect of NAC was measured by DCFH-DA through flow cytometer. The concentration with better antioxidant effect was used for following experiments. (a, b) The level of cytosolic ROS in different groups. The mean fluorescence intensity (MFI) of DCF is used to measure cellular ROS levels. BSA–treated cells without DCFH-DA incubation serve as negative control. (c) Apoptotic cells in different groups. Annexin V positive cells (including Annexin V single positive population and the Annexin V and PI double positive population) representing apoptotic cells were mainly located in the right upper quadrant and right lower quadrant. (d) Average apoptotic rate in the three groups. (e) Insulin secretion of INS-1 cells stimulated with 3.3 mmol/L glucose or 16.7 mmol/L glucose in the different groups. * P<0.05 vs the corresponding control group; # P<0.05 vs the corresponding GA group. (f) Insulin release index (IRI) of each group. The IRI was adopted as an index to estimate the insulin secretion function of INS-1 cells. IRI = insulin concentration of high glucose stimulation (16.7 mmol/L)/insulin concentration of low glucose stimulation (3.3 mmol/L). (g, h) Protein expression of MMP2 in different groups, including both pro-MMP2(∼72 kD) and active MMP2(∼62 kD). (i, j) The gelatinolytic activity of MMP2 in different groups. The white band in gray background was the location of MMP2, band intensity was used to represent the activity of MMP2 in each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 vs the control group; # P<0.05 vs the GA group.

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Sterility, Flow Cytometry, Concentration Assay, Fluorescence, Incubation, Negative Control, Control, Expressing, Activity Assay

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells were transfected with 1.6 µg MMP2 recombination plasmids and 4 µL Lipofectmine 2000 in each well. An empty vector served as control. After 48 h incubation, the mRNA, protein and gelatinolytic activity of MMP2 were evaluated. (a) Relative mRNA level of MMP2 after 48 h transfection in each group. (b, c) Protein expression (including pro-MMP2 and active MMP2) after 48 h of transfection in each group. (d, e) Gelatinolytic activity of MMP2 after 48 h of transfection in each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 vs the control group.

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Transfection, Plasmid Preparation, Control, Incubation, Activity Assay, Expressing

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells were transfected with 1.6 µg MMP2 recombination plasmid and 4 µL Lipofectmine 2000 in each well. An empty vector served as control. The apoptosis and insulin-secretion function of INS-1 cells were evaluated in each group after 48 h of incubation. (a) Apoptotic cells in the control group. Annexin V positive cells (including Annexin V single positive population and the Annexin V and PI double positive population) representing apoptotic cells were mainly located in the right upper quadrant and right lower quadrant. (b) Average apoptotic rate of each group. (c) Insulin secretion stimulated under 3.3 mmol/L or 16.7 mmol/L glucose in each group. * P<0.05 compared to the corresponding control group. (d) Insulin release index (IRI) of each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 vs the control group.

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Transfection, Plasmid Preparation, Control, Incubation

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells were treated with GA (200 mg/L) or BSA (200 mg/L) for 24 h. MMP2 inhibitor (OA-Hy) was dissolved in DMSO and used at a final concentration of 20 µmol/L with GA for 24 h. (a, b) The gelatinolytic activity of MMP2 in different groups. (c) Apoptotic cells in different groups. Annexin V positive cells (including Annexin V single positive population and the Annexin V and PI double positive population) representing apoptotic cells were mainly located in the right upper quadrant and right lower quadrant. (d) Average apoptotic rate in the three groups. (e) Insulin secretion of INS-1 cells stimulated with 3.3 mmol/L glucose or 16.7 mmol/L glucose in the different groups. * P<0.05 vs the corresponding control group; # P<0.05 vs corresponding GA-treated group (f) Insulin release index (IRI) of each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 vs the control group; # P<0.05 vs GA-treated group.

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Concentration Assay, Activity Assay, Control

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells of each well were transfected with 40 pmol MMP-2 siRNA by 2 µL Lipofectmine 2000 in each well. A scrambled siRNA was used as control. 48 h after siRNA transfection, INS-1 cells were then treated with GA (200 mg/L) or BSA (200 mg/L) for another 24 h. (a) mRNA expression of MMP2 in different groups; (b–d) Protein expression (including pro-MMP2 and active MMP2) in each group; (e, f) Gelatinolytic activity of MMP2 in each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 , # P<0.05 vs corresponding control.

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Transfection, Control, Expressing, Activity Assay

Journal: PLoS ONE

Article Title: Matrix Metalloproteinase 2 Contributes to Pancreatic Beta Cell Injury Induced by Oxidative Stress

doi: 10.1371/journal.pone.0110227

Figure Lengend Snippet: INS-1 cells were transfected with 40 pmol MMP-2 siRNA. A scramble siRNA was used as control. 48 h after siRNA transfection, INS-1 cells were then treated with GA (200 mg/L) or BSA (200 mg/L) for another 24 h. (a) Apoptotic cells in different groups. Annexin V positive cells (including Annexin V single positive population and the Annexin V and PI double positive population) representing apoptotic cells were mainly located in the right upper quadrant and right lower quadrant. (b) Average apoptotic rate of each group. (c) The insulin secretion stimulated under 3.3 mmol/L glucose or 16.7 mmol/L glucose in each group. * P<0.05 vs BSA-treated subgroup under 16.7 mmol/L glucose stimulation. # P<0.05 vs corresponding control (treated with GA) under 16.7 mmol/L glucose stimulation. (d) Insulin release index (IRI) of each group. Data are shown as means ± SD, P<0.05 indicates a statistically significant difference. * P<0.05 vs BSA-treated subgroup, # P<0.05 vs corresponding control (treated with GA).

Article Snippet: To determine the protective role of MMP2 inhibition in beta cell dysfunction induced by oxidative stress, INS-1 cells were co-treated with GA and a MMP2 specific inhibitor (cis-9-Octadecenoyl-N- hydroxylamide, OA-Hy, Santa Cruz Biotechnology, Dallas, Texas, USA) for 24 h, the other two groups were treated with GA or BSA.

Techniques: Transfection, Control

Journal: Cell Death & Disease

Article Title: Pleiotropic actions of miR-21 highlight the critical role of deregulated stromal microRNAs during colorectal cancer progression

doi: 10.1038/cddis.2013.213

Figure Lengend Snippet: Ectopic miR-21 expression in MRC5 fibroblasts leads to upregulated MMP2 activity – SW480 CRC cell invasion is MMP2 dependent. ( a ) Representative experiment showing zymographic activity in CM 21 or CM SCC , with and without MMP2 inhibitor (MMP2inh). ( b ) Assessment of MMP2 activity by determination of zymographical band intensity. Results are expressed as a ratio of MMP2 activity in CM 21 compared with CM SCC in three independent experiments. ( c ) Transwell invasion assay using CM 21 or CM SCC as chemoattractant, with and without MMP2 inhibitor. Results represent the mean number of invading cells from four repeat experiments (* P <0.05)

Article Snippet: Specific MMP2 inhibitor I (Merck, Hoddesdon, UK), prepared at a stock concentration of 10 mM in DMSO was also added to the chemoattractant at a final concentration of 25 μ M, which achieved maximal inhibition of invasion with no cytotoxic effects (data not shown).

Techniques: Expressing, Activity Assay, Transwell Invasion Assay

Journal: Cell Death & Disease

Article Title: Pleiotropic actions of miR-21 highlight the critical role of deregulated stromal microRNAs during colorectal cancer progression

doi: 10.1038/cddis.2013.213

Figure Lengend Snippet: Proposed impact of deregulated miR-21 expression in CRC-associated stroma. Reciprocal signalling between stromal and epithelial cells (thin black arrows; left panel) is essential for normal colonic tissue homoeostasis. Early in cancer, this process is corrupted and secreted factors from transformed epithelial cells (dotted black arrow; central panel) initiate profound changes in adjacent stromal cells. Upregulated miR-21 expression in stromal fibroblasts supports TGF- β -dependent fibroblast-to-myofibroblast transdifferentiation (thick black arrow) and causes downregulated RECK expression. Myofibroblast-derived factors are important mediators of tumour progression (dotted red arrow; right panel), and although in the absence of TGF- β , miR-21 is not sufficient to initiate myofibroblast transdifferentiation, upregulated miR-21 does promote tumour proliferation and chemoresistance and enhanced tumour invasion through upregulated MMP2 activity

Article Snippet: Specific MMP2 inhibitor I (Merck, Hoddesdon, UK), prepared at a stock concentration of 10 mM in DMSO was also added to the chemoattractant at a final concentration of 25 μ M, which achieved maximal inhibition of invasion with no cytotoxic effects (data not shown).

Techniques: Expressing, Transformation Assay, Derivative Assay, Activity Assay