coomassie  (Azure Biosystems)


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    Structured Review

    Azure Biosystems coomassie
    Determination of protein expression and PKA-mediated phosphorylation of cMyBPC and other sarcomeric proteins. (a) Representative Western blot and quantification of cMyBPC showing total protein expression. (b) Representative gels shown are stained by Pro-Q (left) for total protein phosphorylation, and the same gel is shown for total protein level (right) stained with <t>Coomassie</t> Blue. Relative protein phosphorylation (phosphorylated signal/total protein signal) was calculated for each protein and is expressed as % of NTG -PKA values for that protein. Values are expressed as mean ± S.E.M., from four hearts in each group. * P
    Coomassie, supplied by Azure Biosystems, used in various techniques. Bioz Stars score: 92/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/coomassie/product/Azure Biosystems
    Average 92 stars, based on 124 article reviews
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    coomassie - by Bioz Stars, 2021-01
    92/100 stars

    Images

    1) Product Images from "The HCM-causing Y235S cMyBPC mutation accelerates contractile function by altering C1 domain structure"

    Article Title: The HCM-causing Y235S cMyBPC mutation accelerates contractile function by altering C1 domain structure

    Journal: Biochimica et biophysica acta. Molecular basis of disease

    doi: 10.1016/j.bbadis.2019.01.007

    Determination of protein expression and PKA-mediated phosphorylation of cMyBPC and other sarcomeric proteins. (a) Representative Western blot and quantification of cMyBPC showing total protein expression. (b) Representative gels shown are stained by Pro-Q (left) for total protein phosphorylation, and the same gel is shown for total protein level (right) stained with Coomassie Blue. Relative protein phosphorylation (phosphorylated signal/total protein signal) was calculated for each protein and is expressed as % of NTG -PKA values for that protein. Values are expressed as mean ± S.E.M., from four hearts in each group. * P
    Figure Legend Snippet: Determination of protein expression and PKA-mediated phosphorylation of cMyBPC and other sarcomeric proteins. (a) Representative Western blot and quantification of cMyBPC showing total protein expression. (b) Representative gels shown are stained by Pro-Q (left) for total protein phosphorylation, and the same gel is shown for total protein level (right) stained with Coomassie Blue. Relative protein phosphorylation (phosphorylated signal/total protein signal) was calculated for each protein and is expressed as % of NTG -PKA values for that protein. Values are expressed as mean ± S.E.M., from four hearts in each group. * P

    Techniques Used: Expressing, Western Blot, Staining

    2) Product Images from "Aquaporin-4-dependent glymphatic solute transport in the rodent brain"

    Article Title: Aquaporin-4-dependent glymphatic solute transport in the rodent brain

    Journal: eLife

    doi: 10.7554/eLife.40070

    Evidence evaluating the role of AQP4 in CSF influx and ISF efflux. ( a ) Meta-analysis from experiments that delivered either fluorescence- or radio-labeled tracers into the cisterna magna of both Aqp4 KO and wild-type rodents. ( b ) Meta-analysis from studies that delivered intracerebral tracers to evaluate clearance or transport of tracers out of the brain. p-Value is from the overall random effects model. Data in forest plots presented as standardized mean difference (SMD) with a 95% confidence interval (CI). Th: thalamus; Cn: caudate nucleus.
    Figure Legend Snippet: Evidence evaluating the role of AQP4 in CSF influx and ISF efflux. ( a ) Meta-analysis from experiments that delivered either fluorescence- or radio-labeled tracers into the cisterna magna of both Aqp4 KO and wild-type rodents. ( b ) Meta-analysis from studies that delivered intracerebral tracers to evaluate clearance or transport of tracers out of the brain. p-Value is from the overall random effects model. Data in forest plots presented as standardized mean difference (SMD) with a 95% confidence interval (CI). Th: thalamus; Cn: caudate nucleus.

    Techniques Used: Fluorescence, Labeling

    Deletion of the adapter protein α-syntrophin impairs AQP4 perivascular localization, and CSF influx into the brain parenchyma. Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) was acquired on a 11.75 preclinical MRI scanner, and was used to characterize the effect of α-syntrophin deletion on gaditeridol influx into the brain. Representative images of AQP4 perivascular localization in wild-type mice ( a ), and the loss of perivascular localization of AQP4 seen in the Snta1 -/- mice ( b ). Scale bar: 50 µm, inset scale bar: 10 µm. ( c-h ) Coronal slice of T 1 -weighted images acquired by DCE-MRI demonstrate the reduced influx of gaditeridol contrast agent into the parenchyma in Snta1 -/- mice relative to wild-type mice at 30 and 60 min. Scale bar: 1 mm. ( i-n ) Quantification of T 1 weighted signal in various brain subregions normalized to baseline at each time point. Traces for each individual animal are presented (lines) along with the summary statistics (mean ±SEM, two-way ANOVA). WT n = 5, ASYNKO n = 7. CTx = cortex (p = 0.0035) Hip = hippocampus (p = 0.0003) Subcortical = subcortical regions (p = 0.0185) 3V = 3 rd Ventricle (p = 0.0284) Total (p = 0.0085) ( Figure 6—source data 1 ). 10.7554/eLife.40070.012 Source data for Figure 6 .
    Figure Legend Snippet: Deletion of the adapter protein α-syntrophin impairs AQP4 perivascular localization, and CSF influx into the brain parenchyma. Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) was acquired on a 11.75 preclinical MRI scanner, and was used to characterize the effect of α-syntrophin deletion on gaditeridol influx into the brain. Representative images of AQP4 perivascular localization in wild-type mice ( a ), and the loss of perivascular localization of AQP4 seen in the Snta1 -/- mice ( b ). Scale bar: 50 µm, inset scale bar: 10 µm. ( c-h ) Coronal slice of T 1 -weighted images acquired by DCE-MRI demonstrate the reduced influx of gaditeridol contrast agent into the parenchyma in Snta1 -/- mice relative to wild-type mice at 30 and 60 min. Scale bar: 1 mm. ( i-n ) Quantification of T 1 weighted signal in various brain subregions normalized to baseline at each time point. Traces for each individual animal are presented (lines) along with the summary statistics (mean ±SEM, two-way ANOVA). WT n = 5, ASYNKO n = 7. CTx = cortex (p = 0.0035) Hip = hippocampus (p = 0.0003) Subcortical = subcortical regions (p = 0.0185) 3V = 3 rd Ventricle (p = 0.0284) Total (p = 0.0085) ( Figure 6—source data 1 ). 10.7554/eLife.40070.012 Source data for Figure 6 .

    Techniques Used: Magnetic Resonance Imaging, Mouse Assay

    NMU: Aqp4 gene deletion reduced the penetration of intracisternally injected tracer into the brain parenchyma. Texas Red-conjugated dextran (TRd3, 3kD) was injected intracisternally into WT and Aqp4 KO mice. Thirty minutes after injection, the anesthetized animals were perfusion fixed, and the fluorescence was evaluated ex vivo. ( a ) Representative near infrared (NIR) fluorescence images of the dorsal and ventral whole-brains of four mice per genotype. ( b-c ) Quantification of the mean integrated optical density (MIOD) of TRd3 on the dorsal (b) and ventral (c) brain surface of WT (grey) and Aqp4 KO (purple) mice from 4.0 mm anterior to 8.0 mm posterior to bregma. ( d ) Representative images of coronal brain sections at +0.5 mm from bregma from five pairs of WT and Aqp4 KO mice showing TRd3 distribution within the brain. ( e ) High magnification micrographs of the hypothalamus (lined area in d) showing the fluorescence intensity of TRd3 within the perivascular space (star) and adjacent brain parenchyma (dotted line) of WT mice and Aqp4 KO mice, respectively. ( f ) Quantification of the percentage area of whole-slice fluorescence of the both genotypes for 6–8 forebrain sections (+1.7 to −0.7 mm from bregma) of each mouse. ( g ) Diagram showing the subregional analysis of brain sections at the level of 0.5 mm anterior to bregma. ( h ) Quantification of the mean fluorescence intensity (AU, arbitrary units) of TRd3 of the dorsal, ventral and lateral brain regions, respectively. ( i-j ) Quantification of the mean fluorescence intensity of TRd3 along the perivascular space and the interstitium adjacent to the vessels under the ventral surface of the hypothalamus of the both genotypes. Shades and error bars represent standard deviation. Data in Figure 2b and c were analyzed by repeated-measures ANOVA, N = 4 per group. Data in Figure 2f,h,i and j were analyzed by Student’s t-test. p-Values shown are comparisons between WT and Aqp4 -/- . ns: not significant ( Figure 2—source data 1 ). 10.7554/eLife.40070.004 Source data for Figure 2 .
    Figure Legend Snippet: NMU: Aqp4 gene deletion reduced the penetration of intracisternally injected tracer into the brain parenchyma. Texas Red-conjugated dextran (TRd3, 3kD) was injected intracisternally into WT and Aqp4 KO mice. Thirty minutes after injection, the anesthetized animals were perfusion fixed, and the fluorescence was evaluated ex vivo. ( a ) Representative near infrared (NIR) fluorescence images of the dorsal and ventral whole-brains of four mice per genotype. ( b-c ) Quantification of the mean integrated optical density (MIOD) of TRd3 on the dorsal (b) and ventral (c) brain surface of WT (grey) and Aqp4 KO (purple) mice from 4.0 mm anterior to 8.0 mm posterior to bregma. ( d ) Representative images of coronal brain sections at +0.5 mm from bregma from five pairs of WT and Aqp4 KO mice showing TRd3 distribution within the brain. ( e ) High magnification micrographs of the hypothalamus (lined area in d) showing the fluorescence intensity of TRd3 within the perivascular space (star) and adjacent brain parenchyma (dotted line) of WT mice and Aqp4 KO mice, respectively. ( f ) Quantification of the percentage area of whole-slice fluorescence of the both genotypes for 6–8 forebrain sections (+1.7 to −0.7 mm from bregma) of each mouse. ( g ) Diagram showing the subregional analysis of brain sections at the level of 0.5 mm anterior to bregma. ( h ) Quantification of the mean fluorescence intensity (AU, arbitrary units) of TRd3 of the dorsal, ventral and lateral brain regions, respectively. ( i-j ) Quantification of the mean fluorescence intensity of TRd3 along the perivascular space and the interstitium adjacent to the vessels under the ventral surface of the hypothalamus of the both genotypes. Shades and error bars represent standard deviation. Data in Figure 2b and c were analyzed by repeated-measures ANOVA, N = 4 per group. Data in Figure 2f,h,i and j were analyzed by Student’s t-test. p-Values shown are comparisons between WT and Aqp4 -/- . ns: not significant ( Figure 2—source data 1 ). 10.7554/eLife.40070.004 Source data for Figure 2 .

    Techniques Used: Injection, Mouse Assay, Fluorescence, Ex Vivo, Standard Deviation

    UNC: CSF tracer influx is decreased in Aqp4 KO mice. ( a ) Coronal sections from a C57BL/6 wild-type mouse (WT), CD1 background strain control ( Aqp4 +/+ ), and Aqp4 KO mice ( Aqp4 -/- ) showing a fluorescent CSF tracer, BSA-647 and co-labeling with DAPI. Scale bar: 1 mm ( b ) Mean pixel intensity in arbitrary units (A.U.) for six brain sections of each mouse for all three groups. n = 3 (WT), 7 ( Aqp4 +/+ ), 6 ( Aqp4 -/- ). One-way ANOVA Tukey’s multiple comparisons test, Interaction term: p = 0.0110, F = 6.512, ns: not significant. ( c ) Diagram showing the anterior-posterior range of the quantified coronal sections relative to bregma from (b). ( d ) Quantification of the slices shown in (c) + 1.2 to −1.8 mm from bregma. Repeated measures two-way ANOVA with Tukey’s multiple comparisons test, Interaction term: p = 0.038, F = 2.085, p values shown are comparisons of WT and Aqp4 +/+ vs. Aqp4 -/- . ( e ) Diagram depicting the ROIs included in the regional analysis of brain slices at +0.6 mm from bregma. CPu: caudoputamen; HT: hypothalamus; BF: basal forebrain; DC: dorsal cortex; LC: lateral cortex; VC: ventral cortex. ( f ) Mean pixel intensity of brain regions shown in (e) for coronal sections + 0.6 mm from bregma. Repeated measures two-way ANOVA Tukey’s multiple comparisons test, Interaction term: p
    Figure Legend Snippet: UNC: CSF tracer influx is decreased in Aqp4 KO mice. ( a ) Coronal sections from a C57BL/6 wild-type mouse (WT), CD1 background strain control ( Aqp4 +/+ ), and Aqp4 KO mice ( Aqp4 -/- ) showing a fluorescent CSF tracer, BSA-647 and co-labeling with DAPI. Scale bar: 1 mm ( b ) Mean pixel intensity in arbitrary units (A.U.) for six brain sections of each mouse for all three groups. n = 3 (WT), 7 ( Aqp4 +/+ ), 6 ( Aqp4 -/- ). One-way ANOVA Tukey’s multiple comparisons test, Interaction term: p = 0.0110, F = 6.512, ns: not significant. ( c ) Diagram showing the anterior-posterior range of the quantified coronal sections relative to bregma from (b). ( d ) Quantification of the slices shown in (c) + 1.2 to −1.8 mm from bregma. Repeated measures two-way ANOVA with Tukey’s multiple comparisons test, Interaction term: p = 0.038, F = 2.085, p values shown are comparisons of WT and Aqp4 +/+ vs. Aqp4 -/- . ( e ) Diagram depicting the ROIs included in the regional analysis of brain slices at +0.6 mm from bregma. CPu: caudoputamen; HT: hypothalamus; BF: basal forebrain; DC: dorsal cortex; LC: lateral cortex; VC: ventral cortex. ( f ) Mean pixel intensity of brain regions shown in (e) for coronal sections + 0.6 mm from bregma. Repeated measures two-way ANOVA Tukey’s multiple comparisons test, Interaction term: p

    Techniques Used: Mouse Assay, Labeling

    Strategy used for generation of KO mice and the experimental design of the study. Top row of each panel represents the institution where the line originated from and the strategy used to generate the four global Aqp4 KO mice and the Snta1 KO mice. Second row of each panel shows the five labs that collected data on glymphatic system function in the five transgenic mouse lines: ( a ) Nanjing Medical University (NMU), ( b ) RIKEN Center for Brain Science (RIKEN), ( c ) University of North Carolina (UNC), ( d ) Oregon Health and Science University (OHSU), ( e ) University of Rochester Medical Center (URMC). ( a-e ) Immunohistochemical analysis showing the lack of AQP4 expression in the global KOs and (d) lack of perivascular AQP4 localization in Snta1 KO compared to WT mice. Scale bar: 50 µm. (a-e) Third row displays the volume and rate used for the intracisterna magna (CM) injections for each experiment, the tracer used, and the experiment duration. Note that URMC collected two full data sets using an injection rate of either 1 or 2 µl/min. ( a-e ) The last row display the analyses strategy employed by each of the five research group in Figures 2 – 6 . ( f ) Additional experiments performed in Figure 7 tested the effect of intrastriatal (IS) injection on global glymphatic function. TxRd, Texas Red; BDA, biotinylated dextran amine; BSA-647, bovine serum albumin-Alexa Fluor 647; DCE-MRI, dynamic contrast-enhanced magnetic resonance imaging.
    Figure Legend Snippet: Strategy used for generation of KO mice and the experimental design of the study. Top row of each panel represents the institution where the line originated from and the strategy used to generate the four global Aqp4 KO mice and the Snta1 KO mice. Second row of each panel shows the five labs that collected data on glymphatic system function in the five transgenic mouse lines: ( a ) Nanjing Medical University (NMU), ( b ) RIKEN Center for Brain Science (RIKEN), ( c ) University of North Carolina (UNC), ( d ) Oregon Health and Science University (OHSU), ( e ) University of Rochester Medical Center (URMC). ( a-e ) Immunohistochemical analysis showing the lack of AQP4 expression in the global KOs and (d) lack of perivascular AQP4 localization in Snta1 KO compared to WT mice. Scale bar: 50 µm. (a-e) Third row displays the volume and rate used for the intracisterna magna (CM) injections for each experiment, the tracer used, and the experiment duration. Note that URMC collected two full data sets using an injection rate of either 1 or 2 µl/min. ( a-e ) The last row display the analyses strategy employed by each of the five research group in Figures 2 – 6 . ( f ) Additional experiments performed in Figure 7 tested the effect of intrastriatal (IS) injection on global glymphatic function. TxRd, Texas Red; BDA, biotinylated dextran amine; BSA-647, bovine serum albumin-Alexa Fluor 647; DCE-MRI, dynamic contrast-enhanced magnetic resonance imaging.

    Techniques Used: Mouse Assay, Transgenic Assay, Immunohistochemistry, Expressing, Injection, Magnetic Resonance Imaging

    RIKEN: Aqp4 -/- mice display compromised CSF tracer infiltration under ketamine-xylazine anesthesia. ( a ) Schematic diagram for CM injection of BDA tracer (left) and experiment schedule (right). ( b ) Examples of SA-enhanced BDA distribution 30 min after CM injection. Slices at an anterior-posterior position of bregma are presented for WT (upper) and Aqp4 -/- (lower) mice. Depth profile is calculated for the cortical position 3 mm lateral to the midline. ( c ) Mean depth profiles of SA-enhanced BDA signals for WT (black, N = 4) and Aqp4 -/- (green, N = 9) mice. ( d ) Mean SA-enhanced BDA signal intensities (3 mm lateral to the midline, depths 0–800 µm) along anterior-posterior positions for WT (black) and Aqp4 -/- (green) mice. Shades and error bars represent SEM. *p
    Figure Legend Snippet: RIKEN: Aqp4 -/- mice display compromised CSF tracer infiltration under ketamine-xylazine anesthesia. ( a ) Schematic diagram for CM injection of BDA tracer (left) and experiment schedule (right). ( b ) Examples of SA-enhanced BDA distribution 30 min after CM injection. Slices at an anterior-posterior position of bregma are presented for WT (upper) and Aqp4 -/- (lower) mice. Depth profile is calculated for the cortical position 3 mm lateral to the midline. ( c ) Mean depth profiles of SA-enhanced BDA signals for WT (black, N = 4) and Aqp4 -/- (green, N = 9) mice. ( d ) Mean SA-enhanced BDA signal intensities (3 mm lateral to the midline, depths 0–800 µm) along anterior-posterior positions for WT (black) and Aqp4 -/- (green) mice. Shades and error bars represent SEM. *p

    Techniques Used: Mouse Assay, Injection

    URMC: Glymphatic influx of CSF tracer is facilitated by AQP4. ( a ) Representative images from an in vivo transcranial optical imaging experiment of a WT control mice ( Aqp4 +/+ ) and Aqp4 KO ( Aqp4 -/- ) mice starting at 15 min after intracisternal delivery of 10 µl (2 µl/min) of a 66 kDa fluorescent tracer, BSA-647. Scale bar: 2 mm. ( b ) Mean pixel intensity (MPI) of BSA-647 over a 30 min experiment, imaging was started at the beginning of the injection. Two-way repeated measures ANOVA with Sidak’s multiple comparisons test, overall model *p = 0.0329, multiple comparisons *p
    Figure Legend Snippet: URMC: Glymphatic influx of CSF tracer is facilitated by AQP4. ( a ) Representative images from an in vivo transcranial optical imaging experiment of a WT control mice ( Aqp4 +/+ ) and Aqp4 KO ( Aqp4 -/- ) mice starting at 15 min after intracisternal delivery of 10 µl (2 µl/min) of a 66 kDa fluorescent tracer, BSA-647. Scale bar: 2 mm. ( b ) Mean pixel intensity (MPI) of BSA-647 over a 30 min experiment, imaging was started at the beginning of the injection. Two-way repeated measures ANOVA with Sidak’s multiple comparisons test, overall model *p = 0.0329, multiple comparisons *p

    Techniques Used: In Vivo, Optical Imaging, Mouse Assay, Imaging, Injection

    3) Product Images from "Poly (ADP-ribose) polymerase 1 mRNA levels strongly correlate with the prognosis of myelodysplastic syndromes"

    Article Title: Poly (ADP-ribose) polymerase 1 mRNA levels strongly correlate with the prognosis of myelodysplastic syndromes

    Journal: Blood Cancer Journal

    doi: 10.1038/bcj.2016.127

    Box plots for the distribution of PARP1 mRNA levels ( a ) in the different types of MDS according to the 2008/2016 WHO classification, ( b ) in the WHO cumulative groups (MDS without excess blasts and MDS with excess blasts), ( c ) in the risk groups according to IPSS, ( d ) in the cumulative risk groups according to IPSS (lower, incorporating low and intermediate-1, and higher, incorporating intermediate-2 and high), ( e ) in the risk groups according to IPSS-R, and ( f ) in the cytogenetic risk groups (per IPSS-R). IPSS, international prognostic scoring system; IPSS-R, revised international prognostic scoring system; MDS, myelodysplastic syndrome; MLD, multilineage dysplasia; RA, refractory anemia; RARS, refractory anemia with ring sideroblasts; RCMD, refractory cytopenia with multilineage dysplasia; RS, ring sideroblasts; SLD, single lineage dysplasia; RAEB, refractory anemia with excess blasts; EB, excess blasts; WHO, world health organization.
    Figure Legend Snippet: Box plots for the distribution of PARP1 mRNA levels ( a ) in the different types of MDS according to the 2008/2016 WHO classification, ( b ) in the WHO cumulative groups (MDS without excess blasts and MDS with excess blasts), ( c ) in the risk groups according to IPSS, ( d ) in the cumulative risk groups according to IPSS (lower, incorporating low and intermediate-1, and higher, incorporating intermediate-2 and high), ( e ) in the risk groups according to IPSS-R, and ( f ) in the cytogenetic risk groups (per IPSS-R). IPSS, international prognostic scoring system; IPSS-R, revised international prognostic scoring system; MDS, myelodysplastic syndrome; MLD, multilineage dysplasia; RA, refractory anemia; RARS, refractory anemia with ring sideroblasts; RCMD, refractory cytopenia with multilineage dysplasia; RS, ring sideroblasts; SLD, single lineage dysplasia; RAEB, refractory anemia with excess blasts; EB, excess blasts; WHO, world health organization.

    Techniques Used:

    Overall survival in relation to PARP1 mRNA levels; OS was evaluated at the cutoff of 0.011, which was estimated to be the best cutoff level of the present series. Results for the whole cohort ( a ), for the subgroup of patients without excess blasts per the 2016 WHO classification ( b ), and for the subgroup of patients with lower risk (low and intermediate-1) per IPSS ( c ). OS, overall survival; PARP1, poly [ADP-ribose] polymerase 1.
    Figure Legend Snippet: Overall survival in relation to PARP1 mRNA levels; OS was evaluated at the cutoff of 0.011, which was estimated to be the best cutoff level of the present series. Results for the whole cohort ( a ), for the subgroup of patients without excess blasts per the 2016 WHO classification ( b ), and for the subgroup of patients with lower risk (low and intermediate-1) per IPSS ( c ). OS, overall survival; PARP1, poly [ADP-ribose] polymerase 1.

    Techniques Used:

    4) Product Images from "Microfluidic rapid and autonomous analytical device (microRAAD) to detect HIV from whole blood samples †"

    Article Title: Microfluidic rapid and autonomous analytical device (microRAAD) to detect HIV from whole blood samples †

    Journal: Lab on a chip

    doi: 10.1039/c9lc00506d

    Detection of HIV RNA and virus amplified by RT-LAMP. Electrophoresis gels verifying amplification (top, contrast increased for visualization), LFIA test results (middle), and LFIA test line quantification (bottom). (A) Labeled RT-LAMP amplification products are visually detectable from as few as 10 copies of HIV RNA diluted in water. (B) Labeled RT-LAMP amplification products are visually detectable from as few as 10,000 HIV viral particles when reactions contain 16% serum. n=3, replicates indicated by each circle; *** indicates p ≤ 0.001; ** indicates p ≤ 0.01; * indicates p ≤ 0.05.
    Figure Legend Snippet: Detection of HIV RNA and virus amplified by RT-LAMP. Electrophoresis gels verifying amplification (top, contrast increased for visualization), LFIA test results (middle), and LFIA test line quantification (bottom). (A) Labeled RT-LAMP amplification products are visually detectable from as few as 10 copies of HIV RNA diluted in water. (B) Labeled RT-LAMP amplification products are visually detectable from as few as 10,000 HIV viral particles when reactions contain 16% serum. n=3, replicates indicated by each circle; *** indicates p ≤ 0.001; ** indicates p ≤ 0.01; * indicates p ≤ 0.05.

    Techniques Used: Amplification, Electrophoresis, Labeling

    Detection of HIV virus diluted in whole blood on microRAAD with reagents dried for 21 days. (A) Representative μPADs imaged 90 minutes after blood (with and without HIV virus) deposited into microRAAD's sample inlet. After capillary migration of HIV from sample inlet to amplification zone and subsequent heating, the valves are automatically heated, releasing solution to LFIA for detection. As few as 3 × 10 5 HIV (B) osmotically lysed virus copies in rehydrating mix alone or (C) intact virus in rehydrating mix with 12 μL of blood are detectable by microRAAD prepared with RT-LAMP reagents dried for 21 days. n = 4 (B) and n = 3 (C), replicates indicated by each circle; ** indicates p ≤ 0.05 and ** indicates p ≤ 0.01.
    Figure Legend Snippet: Detection of HIV virus diluted in whole blood on microRAAD with reagents dried for 21 days. (A) Representative μPADs imaged 90 minutes after blood (with and without HIV virus) deposited into microRAAD's sample inlet. After capillary migration of HIV from sample inlet to amplification zone and subsequent heating, the valves are automatically heated, releasing solution to LFIA for detection. As few as 3 × 10 5 HIV (B) osmotically lysed virus copies in rehydrating mix alone or (C) intact virus in rehydrating mix with 12 μL of blood are detectable by microRAAD prepared with RT-LAMP reagents dried for 21 days. n = 4 (B) and n = 3 (C), replicates indicated by each circle; ** indicates p ≤ 0.05 and ** indicates p ≤ 0.01.

    Techniques Used: Migration, Amplification

    Detection of HIV virus amplified by dried RT-LAMP reagents. (A) There is no significant difference in test line intensity of labeled amplification products detected on LFIAs after amplification with fresh RT-LAMP reagents as with reagents dried for 21 days. n=5 (fresh), n=13 (dried), circles indicate replicates; *** indicates p ≤ 0.001 (B) Labeled RT-LAMP amplification products are visually detectable from as few as 1,000 HIV virus particles when reactions contain 16% serum. Electrophoresis gels verifying amplification (top, contrast increased for visualization), LFIA test results (middle), and LFIA test line quantification (bottom). n=3, circles indicate replicates; ** indicates p ≤ 0.01; * indicates p ≤ 0.05.
    Figure Legend Snippet: Detection of HIV virus amplified by dried RT-LAMP reagents. (A) There is no significant difference in test line intensity of labeled amplification products detected on LFIAs after amplification with fresh RT-LAMP reagents as with reagents dried for 21 days. n=5 (fresh), n=13 (dried), circles indicate replicates; *** indicates p ≤ 0.001 (B) Labeled RT-LAMP amplification products are visually detectable from as few as 1,000 HIV virus particles when reactions contain 16% serum. Electrophoresis gels verifying amplification (top, contrast increased for visualization), LFIA test results (middle), and LFIA test line quantification (bottom). n=3, circles indicate replicates; ** indicates p ≤ 0.01; * indicates p ≤ 0.05.

    Techniques Used: Amplification, Labeling, Electrophoresis

    Detection of HIV virus from spiked blood on microRAAD with reagents dried for 21 days. (A) Representative μPADs imaged 90 minutes after blood with and without HIV virus deposited into microRAAD’s sample inlets. After capillary migration of HIV to RT-LAMP zone, the RT-LAMP zone and valves are automatically heated, releasing solution to LFIA for detection. As few as 105 HIV virus copies in (B) rehydrating mix alone or (C) rehydrating mix with 12μL of blood are detectable by microRAAD prepared with RT-LAMP reagents dried for 21 days. n=4 (B) and n=3 (C), replicates indicated by each circle; ** indicates p ≤ 0.05 and ** indicates p ≤ 0.01.
    Figure Legend Snippet: Detection of HIV virus from spiked blood on microRAAD with reagents dried for 21 days. (A) Representative μPADs imaged 90 minutes after blood with and without HIV virus deposited into microRAAD’s sample inlets. After capillary migration of HIV to RT-LAMP zone, the RT-LAMP zone and valves are automatically heated, releasing solution to LFIA for detection. As few as 105 HIV virus copies in (B) rehydrating mix alone or (C) rehydrating mix with 12μL of blood are detectable by microRAAD prepared with RT-LAMP reagents dried for 21 days. n=4 (B) and n=3 (C), replicates indicated by each circle; ** indicates p ≤ 0.05 and ** indicates p ≤ 0.01.

    Techniques Used: Migration

    5) Product Images from "Homodimerisation-independent cleavage of dsRNA by a pestiviral nicking endoribonuclease"

    Article Title: Homodimerisation-independent cleavage of dsRNA by a pestiviral nicking endoribonuclease

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-26557-4

    Extended E rns substrate specificity. Various dilutions of Strep-tag purified wild-type (C171), monomeric (R171) and RNase-inactive mutant (H30F) E rns were incubated with 625 nM of poly(U), poly(A), poly(C) and poly(G), either labeled in green or red (panels a–d) as described in Table 1 . Samples were separated by 14% PAGE and fluorescence was analysed with an Azure c600 imaging system. Due to the known, defined length of the directly labeled fragments, no size ladder was applied. Non-cropped gels as representative experiment out of three are shown.
    Figure Legend Snippet: Extended E rns substrate specificity. Various dilutions of Strep-tag purified wild-type (C171), monomeric (R171) and RNase-inactive mutant (H30F) E rns were incubated with 625 nM of poly(U), poly(A), poly(C) and poly(G), either labeled in green or red (panels a–d) as described in Table 1 . Samples were separated by 14% PAGE and fluorescence was analysed with an Azure c600 imaging system. Due to the known, defined length of the directly labeled fragments, no size ladder was applied. Non-cropped gels as representative experiment out of three are shown.

    Techniques Used: Strep-tag, Purification, Mutagenesis, Incubation, Labeling, Polyacrylamide Gel Electrophoresis, Fluorescence, Imaging

    6) Product Images from "Epstein-Barr Virus Latent Membrane Protein 1 Regulates Host B Cell MicroRNA-155 and Its Target FOXO3a via PI3K p110α Activation"

    Article Title: Epstein-Barr Virus Latent Membrane Protein 1 Regulates Host B Cell MicroRNA-155 and Its Target FOXO3a via PI3K p110α Activation

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2019.02692

    miR-155 targets FOXO3a and PI3K p85α are downregulated in EBV+ B cell lymphomas. Lysates from EBV+ and EBV− B cell lymphomas were generated with RIPA buffer supplemented with 1× Halt Protease and Phosphatase Inhibitors and 1 mM sodium orthovanadate and quantified using the Pierce 660 nm Protein Assay. About 30 μg of protein was loaded on a 4–20% tris-glycine gel and subsequently transferred to a nitrocellulose membrane. Western blots for the indicated proteins were performed as per manufacturer’s instructions and imaged via the Azure c300 digital imager. Representative blots are shown in (A) . Densitometry was performed using ImageJ, and all values were background subtracted and normalized to the indicated loading control (B–E) . Each point represents an experimental replicate. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001 by linear mixed effects model.
    Figure Legend Snippet: miR-155 targets FOXO3a and PI3K p85α are downregulated in EBV+ B cell lymphomas. Lysates from EBV+ and EBV− B cell lymphomas were generated with RIPA buffer supplemented with 1× Halt Protease and Phosphatase Inhibitors and 1 mM sodium orthovanadate and quantified using the Pierce 660 nm Protein Assay. About 30 μg of protein was loaded on a 4–20% tris-glycine gel and subsequently transferred to a nitrocellulose membrane. Western blots for the indicated proteins were performed as per manufacturer’s instructions and imaged via the Azure c300 digital imager. Representative blots are shown in (A) . Densitometry was performed using ImageJ, and all values were background subtracted and normalized to the indicated loading control (B–E) . Each point represents an experimental replicate. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001 by linear mixed effects model.

    Techniques Used: Generated, Western Blot

    7) Product Images from "Particle Diffusometry: An Optical Detection Method for Vibrio cholerae Presence in Environmental Water Samples"

    Article Title: Particle Diffusometry: An Optical Detection Method for Vibrio cholerae Presence in Environmental Water Samples

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-38056-7

    Detection of V . cholerae amplification using purified DNA. NTC represents no added V . cholerae DNA. ( A ) Real-time fluorescence was monitored over a 20-minute LAMP reaction for initial DNA concentrations between 10 0 –10 5 DNA copies/reaction and ( B ) the corresponding C T values were recorded for each reaction and are not available (NA) for samples that did not amplify. ( C ) A 2% agarose gel confirms amplification and presents the DNA banding pattern of LAMP amplicons at the different dilutions. ( D ) Box plots of the average change in fluorescence ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\rm{\Delta }}EvaGreen/Rox$$\end{document} Δ E v a G r e e n / R o x ) shows a trend of a greater change in fluorescence signal at higher initial V . cholerae DNA concentrations with statistical differences for samples 10 2 (***p-value
    Figure Legend Snippet: Detection of V . cholerae amplification using purified DNA. NTC represents no added V . cholerae DNA. ( A ) Real-time fluorescence was monitored over a 20-minute LAMP reaction for initial DNA concentrations between 10 0 –10 5 DNA copies/reaction and ( B ) the corresponding C T values were recorded for each reaction and are not available (NA) for samples that did not amplify. ( C ) A 2% agarose gel confirms amplification and presents the DNA banding pattern of LAMP amplicons at the different dilutions. ( D ) Box plots of the average change in fluorescence ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\rm{\Delta }}EvaGreen/Rox$$\end{document} Δ E v a G r e e n / R o x ) shows a trend of a greater change in fluorescence signal at higher initial V . cholerae DNA concentrations with statistical differences for samples 10 2 (***p-value

    Techniques Used: Amplification, Purification, Fluorescence, Agarose Gel Electrophoresis

    Illustration of PD-LAMP set-up. ( A ) The LAMP assay is performed in the presence of V . cholerae DNA (left). LAMP amplicons combined with polystyrene fluorescent particles (middle) are imaged under fluorescence microscopy (right). ( B ) Relationship of particle motion and viscosity. Particles undergo Brownian motion in a solution (left). In the presence of LAMP amplicons, the viscosity of the solution increases and particles experience hindered motion, indicating the presence of V . cholerae DNA in the sample (right).
    Figure Legend Snippet: Illustration of PD-LAMP set-up. ( A ) The LAMP assay is performed in the presence of V . cholerae DNA (left). LAMP amplicons combined with polystyrene fluorescent particles (middle) are imaged under fluorescence microscopy (right). ( B ) Relationship of particle motion and viscosity. Particles undergo Brownian motion in a solution (left). In the presence of LAMP amplicons, the viscosity of the solution increases and particles experience hindered motion, indicating the presence of V . cholerae DNA in the sample (right).

    Techniques Used: Lamp Assay, Fluorescence, Microscopy

    8) Product Images from "Aquaporin-4-dependent glymphatic solute transport in the rodent brain"

    Article Title: Aquaporin-4-dependent glymphatic solute transport in the rodent brain

    Journal: eLife

    doi: 10.7554/eLife.40070

    Evidence evaluating the role of AQP4 in CSF influx and ISF efflux. ( a ) Meta-analysis from experiments that delivered either fluorescence- or radio-labeled tracers into the cisterna magna of both Aqp4 KO and wild-type rodents. ( b ) Meta-analysis from studies that delivered intracerebral tracers to evaluate clearance or transport of tracers out of the brain. p-Value is from the overall random effects model. Data in forest plots presented as standardized mean difference (SMD) with a 95% confidence interval (CI). Th: thalamus; Cn: caudate nucleus.
    Figure Legend Snippet: Evidence evaluating the role of AQP4 in CSF influx and ISF efflux. ( a ) Meta-analysis from experiments that delivered either fluorescence- or radio-labeled tracers into the cisterna magna of both Aqp4 KO and wild-type rodents. ( b ) Meta-analysis from studies that delivered intracerebral tracers to evaluate clearance or transport of tracers out of the brain. p-Value is from the overall random effects model. Data in forest plots presented as standardized mean difference (SMD) with a 95% confidence interval (CI). Th: thalamus; Cn: caudate nucleus.

    Techniques Used: Fluorescence, Labeling

    URMC: Glymphatic influx of CSF tracer is facilitated by AQP4. ( a ) Representative images from an in vivo transcranial optical imaging experiment of a WT control mice ( Aqp4 +/+ ) and Aqp4 KO ( Aqp4 -/- ) mice starting at 15 min after intracisternal delivery of 10 µl (2 µl/min) of a 66 kDa fluorescent tracer, BSA-647. Scale bar: 2 mm. ( b ) Mean pixel intensity (MPI) of BSA-647 over a 30 min experiment, imaging was started at the beginning of the injection. Two-way repeated measures ANOVA with Sidak’s multiple comparisons test, overall model *p = 0.0329, multiple comparisons *p
    Figure Legend Snippet: URMC: Glymphatic influx of CSF tracer is facilitated by AQP4. ( a ) Representative images from an in vivo transcranial optical imaging experiment of a WT control mice ( Aqp4 +/+ ) and Aqp4 KO ( Aqp4 -/- ) mice starting at 15 min after intracisternal delivery of 10 µl (2 µl/min) of a 66 kDa fluorescent tracer, BSA-647. Scale bar: 2 mm. ( b ) Mean pixel intensity (MPI) of BSA-647 over a 30 min experiment, imaging was started at the beginning of the injection. Two-way repeated measures ANOVA with Sidak’s multiple comparisons test, overall model *p = 0.0329, multiple comparisons *p

    Techniques Used: In Vivo, Optical Imaging, Mouse Assay, Imaging, Injection

    RIKEN: Aqp4 -/- mice display compromised CSF tracer infiltration under ketamine-xylazine anesthesia. ( a ) Schematic diagram for CM injection of BDA tracer (left) and experiment schedule (right). ( b ) Examples of SA-enhanced BDA distribution 30 min after CM injection. Slices at an anterior-posterior position of bregma are presented for WT (upper) and Aqp4 -/- (lower) mice. Depth profile is calculated for the cortical position 3 mm lateral to the midline. ( c ) Mean depth profiles of SA-enhanced BDA signals for WT (black, N = 4) and Aqp4 -/- (green, N = 9) mice. ( d ) Mean SA-enhanced BDA signal intensities (3 mm lateral to the midline, depths 0–800 µm) along anterior-posterior positions for WT (black) and Aqp4 -/- ).
    Figure Legend Snippet: RIKEN: Aqp4 -/- mice display compromised CSF tracer infiltration under ketamine-xylazine anesthesia. ( a ) Schematic diagram for CM injection of BDA tracer (left) and experiment schedule (right). ( b ) Examples of SA-enhanced BDA distribution 30 min after CM injection. Slices at an anterior-posterior position of bregma are presented for WT (upper) and Aqp4 -/- (lower) mice. Depth profile is calculated for the cortical position 3 mm lateral to the midline. ( c ) Mean depth profiles of SA-enhanced BDA signals for WT (black, N = 4) and Aqp4 -/- (green, N = 9) mice. ( d ) Mean SA-enhanced BDA signal intensities (3 mm lateral to the midline, depths 0–800 µm) along anterior-posterior positions for WT (black) and Aqp4 -/- ).

    Techniques Used: Mouse Assay, Injection

    UNC: CSF tracer influx is decreased in Aqp4 KO mice. ( a ) Coronal sections from a C57BL/6 wild-type mouse (WT), CD1 background strain control ( Aqp4 +/+ ), and Aqp4 KO mice ( Aqp4 -/- ) showing a fluorescent CSF tracer, BSA-647 and co-labeling with DAPI. Scale bar: 1 mm ( b ) Mean pixel intensity in arbitrary units (A.U.) for six brain sections of each mouse for all three groups. n = 3 (WT), 7 ( Aqp4 +/+ ), 6 ( Aqp4 -/- ). One-way ANOVA Tukey’s multiple comparisons test, Interaction term: p = 0.0110, F = 6.512, ns: not significant. ( c ) Diagram showing the anterior-posterior range of the quantified coronal sections relative to bregma from (b). ( d ) Quantification of the slices shown in (c) + 1.2 to −1.8 mm from bregma. Repeated measures two-way ANOVA with Tukey’s multiple comparisons test, Interaction term: p = 0.038, F = 2.085, p values shown are comparisons of WT and Aqp4 +/+ vs. Aqp4 -/- . ( e ) Diagram depicting the ROIs included in the regional analysis of brain slices at +0.6 mm from bregma. CPu: caudoputamen; HT: hypothalamus; BF: basal forebrain; DC: dorsal cortex; LC: lateral cortex; VC: ventral cortex. ( f ).
    Figure Legend Snippet: UNC: CSF tracer influx is decreased in Aqp4 KO mice. ( a ) Coronal sections from a C57BL/6 wild-type mouse (WT), CD1 background strain control ( Aqp4 +/+ ), and Aqp4 KO mice ( Aqp4 -/- ) showing a fluorescent CSF tracer, BSA-647 and co-labeling with DAPI. Scale bar: 1 mm ( b ) Mean pixel intensity in arbitrary units (A.U.) for six brain sections of each mouse for all three groups. n = 3 (WT), 7 ( Aqp4 +/+ ), 6 ( Aqp4 -/- ). One-way ANOVA Tukey’s multiple comparisons test, Interaction term: p = 0.0110, F = 6.512, ns: not significant. ( c ) Diagram showing the anterior-posterior range of the quantified coronal sections relative to bregma from (b). ( d ) Quantification of the slices shown in (c) + 1.2 to −1.8 mm from bregma. Repeated measures two-way ANOVA with Tukey’s multiple comparisons test, Interaction term: p = 0.038, F = 2.085, p values shown are comparisons of WT and Aqp4 +/+ vs. Aqp4 -/- . ( e ) Diagram depicting the ROIs included in the regional analysis of brain slices at +0.6 mm from bregma. CPu: caudoputamen; HT: hypothalamus; BF: basal forebrain; DC: dorsal cortex; LC: lateral cortex; VC: ventral cortex. ( f ).

    Techniques Used: Mouse Assay, Labeling

    Deletion of the adapter protein α-syntrophin impairs AQP4 perivascular localization, and CSF influx into the brain parenchyma. Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) was acquired on a 11.75 preclinical MRI scanner, and was used to characterize the effect of α-syntrophin deletion on gaditeridol influx into the brain. Representative images of AQP4 perivascular localization in wild-type mice ( a ), and the loss of perivascular localization of AQP4 seen in the Snta1 -/- mice ( b ). Scale bar: 50 µm, inset scale bar: 10 µm. ( c-h ) Coronal slice of T 1 -weighted images acquired by DCE-MRI demonstrate the reduced influx of gaditeridol contrast agent into the parenchyma in Snta1 -/- mice relative to wild-type mice at 30 and 60 min. Scale bar: 1 mm. ( i-n ) Quantification of T 1 weighted signal in various brain subregions normalized to baseline at each time point. Traces for each individual animal are presented (lines) along with the summary statistics (mean ±SEM, two-way ANOVA). WT n = 5, ASYNKO n = 7. CTx = cortex (p = 0.0035) Hip = hippocampus (p = 0.0003) Subcortical = subcortical regions (p = 0.0185) 3V = 3 rd ).
    Figure Legend Snippet: Deletion of the adapter protein α-syntrophin impairs AQP4 perivascular localization, and CSF influx into the brain parenchyma. Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) was acquired on a 11.75 preclinical MRI scanner, and was used to characterize the effect of α-syntrophin deletion on gaditeridol influx into the brain. Representative images of AQP4 perivascular localization in wild-type mice ( a ), and the loss of perivascular localization of AQP4 seen in the Snta1 -/- mice ( b ). Scale bar: 50 µm, inset scale bar: 10 µm. ( c-h ) Coronal slice of T 1 -weighted images acquired by DCE-MRI demonstrate the reduced influx of gaditeridol contrast agent into the parenchyma in Snta1 -/- mice relative to wild-type mice at 30 and 60 min. Scale bar: 1 mm. ( i-n ) Quantification of T 1 weighted signal in various brain subregions normalized to baseline at each time point. Traces for each individual animal are presented (lines) along with the summary statistics (mean ±SEM, two-way ANOVA). WT n = 5, ASYNKO n = 7. CTx = cortex (p = 0.0035) Hip = hippocampus (p = 0.0003) Subcortical = subcortical regions (p = 0.0185) 3V = 3 rd ).

    Techniques Used: Magnetic Resonance Imaging, Mouse Assay