Structured Review

Cell Signaling Technology Inc unc5b
Endothelial cells derived from human umbilical vein endothelial cells (HUVEC) express low levels of classic Netrin receptors and both Netrin-1 (NTN-1) and Netrin-4 (NTN-4) ligands. a mRNA levels of classic (■) and non-classic (□) Netrin receptors were quantified by qPCR relative to GAPDH expression ( dashed line ). Values are mean ± S.E.M. ( n = 7). b Absence of DCC expression in HUVEC was established by Western blot, while Neogenin-1 as well as <t>UNC5b</t> and UNC5c expression could be detected. Endogenous Netrins and vascular endothelial growth factor (VEGF) expression in HUVEC was determined by Western blot. β-actin was used as internal reference ( n = 5–7). c The non-classical Netrin receptor integrin α3β1 was also detected by flow cytometry ( n = 3). d Immunofluorescence for receptors and ligands in HUVEC (magnification × 63). eNOS (endothelial nitric oxide synthase) or CD-31 (PECAM) were used as endothelial cell markers and DAPI for nuclear counterstain
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Images

1) Product Images from "Netrin-1 acts as a non-canonical angiogenic factor produced by human Wharton’s jelly mesenchymal stem cells (WJ-MSC)"

Article Title: Netrin-1 acts as a non-canonical angiogenic factor produced by human Wharton’s jelly mesenchymal stem cells (WJ-MSC)

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-017-0494-5

Endothelial cells derived from human umbilical vein endothelial cells (HUVEC) express low levels of classic Netrin receptors and both Netrin-1 (NTN-1) and Netrin-4 (NTN-4) ligands. a mRNA levels of classic (■) and non-classic (□) Netrin receptors were quantified by qPCR relative to GAPDH expression ( dashed line ). Values are mean ± S.E.M. ( n = 7). b Absence of DCC expression in HUVEC was established by Western blot, while Neogenin-1 as well as UNC5b and UNC5c expression could be detected. Endogenous Netrins and vascular endothelial growth factor (VEGF) expression in HUVEC was determined by Western blot. β-actin was used as internal reference ( n = 5–7). c The non-classical Netrin receptor integrin α3β1 was also detected by flow cytometry ( n = 3). d Immunofluorescence for receptors and ligands in HUVEC (magnification × 63). eNOS (endothelial nitric oxide synthase) or CD-31 (PECAM) were used as endothelial cell markers and DAPI for nuclear counterstain
Figure Legend Snippet: Endothelial cells derived from human umbilical vein endothelial cells (HUVEC) express low levels of classic Netrin receptors and both Netrin-1 (NTN-1) and Netrin-4 (NTN-4) ligands. a mRNA levels of classic (■) and non-classic (□) Netrin receptors were quantified by qPCR relative to GAPDH expression ( dashed line ). Values are mean ± S.E.M. ( n = 7). b Absence of DCC expression in HUVEC was established by Western blot, while Neogenin-1 as well as UNC5b and UNC5c expression could be detected. Endogenous Netrins and vascular endothelial growth factor (VEGF) expression in HUVEC was determined by Western blot. β-actin was used as internal reference ( n = 5–7). c The non-classical Netrin receptor integrin α3β1 was also detected by flow cytometry ( n = 3). d Immunofluorescence for receptors and ligands in HUVEC (magnification × 63). eNOS (endothelial nitric oxide synthase) or CD-31 (PECAM) were used as endothelial cell markers and DAPI for nuclear counterstain

Techniques Used: Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Droplet Countercurrent Chromatography, Western Blot, Flow Cytometry, Cytometry, Immunofluorescence

2) Product Images from "Postsynaptic density protein 95 (PSD-95) is transported by KIF5 to dendritic regions"

Article Title: Postsynaptic density protein 95 (PSD-95) is transported by KIF5 to dendritic regions

Journal: Molecular Brain

doi: 10.1186/s13041-019-0520-x

ADPDZ3 expression reduces the level of PSD-95 in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using monoclonal anti-PSD-antibody and Cy3-conjugated anti-mouse IgG antibody or polyclonal anti-synapsin I antibody and Alexa Fluor® 647 anti-rabbit IgG antibody. They were then visualized using confocal microscopy. a Representative images of expressed neurons. Arrows indicate analyzed dendrites. Scale bar: 20 μm. b , c ADPDZ3 expression reduced the number of PSD-95 particles in the dendrites (GFP: 100.00% ± 6.08%, n = 32, 1766 μm; ADPDZ3: 82.27% ± 4.03%, n = 35, 1973 μm; * P
Figure Legend Snippet: ADPDZ3 expression reduces the level of PSD-95 in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using monoclonal anti-PSD-antibody and Cy3-conjugated anti-mouse IgG antibody or polyclonal anti-synapsin I antibody and Alexa Fluor® 647 anti-rabbit IgG antibody. They were then visualized using confocal microscopy. a Representative images of expressed neurons. Arrows indicate analyzed dendrites. Scale bar: 20 μm. b , c ADPDZ3 expression reduced the number of PSD-95 particles in the dendrites (GFP: 100.00% ± 6.08%, n = 32, 1766 μm; ADPDZ3: 82.27% ± 4.03%, n = 35, 1973 μm; * P

Techniques Used: Expressing, Cell Culture, Infection, Incubation, Immunostaining, Confocal Microscopy

ADPDZ3 expression reduces surface glutamate receptor 1 (GluA1) in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using polyclonal GluA1 antibody and Cy3-conjugated anti-mouse IgG antibody. They were visualized using confocal microscopy. a , b Representative images of expressed neurons and selected dendrites in the analyses. Dotted boxes indicate analyzed dendrites. Scale bar: 20 μm. c , d ADPDZ3 expression reduced the number of surface GluA1 particles (GFP: 100.00% ± 7.83%, n = 18, 1486 μm; ADPDZ3: 66.50% ± 6.21%, n = 14, 1254 μm; ** P
Figure Legend Snippet: ADPDZ3 expression reduces surface glutamate receptor 1 (GluA1) in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using polyclonal GluA1 antibody and Cy3-conjugated anti-mouse IgG antibody. They were visualized using confocal microscopy. a , b Representative images of expressed neurons and selected dendrites in the analyses. Dotted boxes indicate analyzed dendrites. Scale bar: 20 μm. c , d ADPDZ3 expression reduced the number of surface GluA1 particles (GFP: 100.00% ± 7.83%, n = 18, 1486 μm; ADPDZ3: 66.50% ± 6.21%, n = 14, 1254 μm; ** P

Techniques Used: Expressing, Cell Culture, Infection, Incubation, Immunostaining, Confocal Microscopy

Expression of KIF5A ΔMD mutant reduces the number and average size of PSD-95 particles in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP, GFP-KIF5A (WT), or GFP-KIF5A (ΔMD), incubated for 9 h, and immunostained with monoclonal anti-PSD-95 antibody. a Representative images of infected neurons. Scale bar: 20 μm. b ΔMD expression significantly reduces the number of PSD-95 particles (GFP: 100.00% ± 3.88%, n = 31 dendrites, 3417 μm; KIF5A [WT]: 111.70% ± 4.91%, n = 31 dendrites, 3724 μm; KIF5A [ΔMD]: 78.48% ± 4.23%, n = 29 dendrites, 3287 μm; Kruskal–Wallis test: P
Figure Legend Snippet: Expression of KIF5A ΔMD mutant reduces the number and average size of PSD-95 particles in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP, GFP-KIF5A (WT), or GFP-KIF5A (ΔMD), incubated for 9 h, and immunostained with monoclonal anti-PSD-95 antibody. a Representative images of infected neurons. Scale bar: 20 μm. b ΔMD expression significantly reduces the number of PSD-95 particles (GFP: 100.00% ± 3.88%, n = 31 dendrites, 3417 μm; KIF5A [WT]: 111.70% ± 4.91%, n = 31 dendrites, 3724 μm; KIF5A [ΔMD]: 78.48% ± 4.23%, n = 29 dendrites, 3287 μm; Kruskal–Wallis test: P

Techniques Used: Expressing, Mutagenesis, Cell Culture, Infection, Incubation

PSD-95 is colocalized with KIF5. a Representative images of immunostaining. Scale bar: 20 μm. Cultured neurons were immunostained with polyclonal anti-PSD-95 antibody and monoclonal anti-KIF5 antibody (H2), followed by Alexa Fluor® 488 anti-rabbit IgG antibody for PSD-95 and Cy3-conjugated anti-mouse IgG antibody for KIF5. b Results of co-localization analysis. 53.17% ± 3.86% ( n = 19 dendrites, 1354 μm) of KIF5-immunopositve puncta are colocalized with PSD-95-immunopositve puncta and 62.55% ± 1.69% (n = 19 dendrites, 1354 μm) of PSD-95-immunopositive puncta are colocalized with KIF5-immunopositve puncta. c Results of proximity ligation assay. Cultured neurons were infected with Sindbis viruses encoding GFP and subjected to PLA. Red dots in PLA indicate an interaction between the two proteins. Scale bar: 20 μm. d Results of IP assays using rat brain lysates. In total 500 μg of rat brain lysate was used in the IP assays using 3 μg monoclonal anti-KIF5 antibodies or polyclonal PSD-95 antibodies. These were analyzed by Western blotting using the antibody indicated. Asterisks indicate interaction bands in the Western blot assays
Figure Legend Snippet: PSD-95 is colocalized with KIF5. a Representative images of immunostaining. Scale bar: 20 μm. Cultured neurons were immunostained with polyclonal anti-PSD-95 antibody and monoclonal anti-KIF5 antibody (H2), followed by Alexa Fluor® 488 anti-rabbit IgG antibody for PSD-95 and Cy3-conjugated anti-mouse IgG antibody for KIF5. b Results of co-localization analysis. 53.17% ± 3.86% ( n = 19 dendrites, 1354 μm) of KIF5-immunopositve puncta are colocalized with PSD-95-immunopositve puncta and 62.55% ± 1.69% (n = 19 dendrites, 1354 μm) of PSD-95-immunopositive puncta are colocalized with KIF5-immunopositve puncta. c Results of proximity ligation assay. Cultured neurons were infected with Sindbis viruses encoding GFP and subjected to PLA. Red dots in PLA indicate an interaction between the two proteins. Scale bar: 20 μm. d Results of IP assays using rat brain lysates. In total 500 μg of rat brain lysate was used in the IP assays using 3 μg monoclonal anti-KIF5 antibodies or polyclonal PSD-95 antibodies. These were analyzed by Western blotting using the antibody indicated. Asterisks indicate interaction bands in the Western blot assays

Techniques Used: Immunostaining, Cell Culture, Proximity Ligation Assay, Infection, Western Blot

3) Product Images from "Novel elucidation and treatment of pancreatic chronic graft-versus-host disease in mice"

Article Title: Novel elucidation and treatment of pancreatic chronic graft-versus-host disease in mice

Journal: Royal Society Open Science

doi: 10.1098/rsos.181067

EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in cGVHD-affected pancreas and its syngeneic control counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane1: syngeneic control subject, lane 2: cGVHD-affected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S9. ( c ) The corresponding quantitative analysis of the protein bands was conducted. cGVHD-affected pancreas (red) and its syngeneic control counterpart (blue). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., control: n = 5, cGVHD: n = 5.
Figure Legend Snippet: EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in cGVHD-affected pancreas and its syngeneic control counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane1: syngeneic control subject, lane 2: cGVHD-affected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S9. ( c ) The corresponding quantitative analysis of the protein bands was conducted. cGVHD-affected pancreas (red) and its syngeneic control counterpart (blue). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., control: n = 5, cGVHD: n = 5.

Techniques Used: Immunostaining, Staining, Fluorescence

EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in the PBA-treated pancreas and its vehicle-treated counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane 1: PBA-injected pancreas, lane 2: vehicle-injected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S19. ( c ) The corresponding quantitative analysis of the protein bands was conducted. PBA-medicated pancreas (blue) and its vehicle-medicated counterpart (red). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., PBA: n = 6, vehicle: n = 6.
Figure Legend Snippet: EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in the PBA-treated pancreas and its vehicle-treated counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane 1: PBA-injected pancreas, lane 2: vehicle-injected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S19. ( c ) The corresponding quantitative analysis of the protein bands was conducted. PBA-medicated pancreas (blue) and its vehicle-medicated counterpart (red). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., PBA: n = 6, vehicle: n = 6.

Techniques Used: Immunostaining, Staining, Fluorescence, Injection

4) Product Images from "IL-8 secretion in primary cultures of prostate cells is associated with prostate cancer aggressiveness"

Article Title: IL-8 secretion in primary cultures of prostate cells is associated with prostate cancer aggressiveness

Journal: Research and Reports in Urology

doi: 10.2147/RRU.S58643

Characterization of human primary prostate epithelial cell cultures. Notes: ( A ) Primary outgrowth of epithelial cells derived from a biopsy specimen. ( B ) Primary epithelial cell colony. ( C ) Characterization of epithelial cells: green fluorescence shows high-molecular-weight cytokeratins; blue fluorescence shows nuclei reactive with 4′,6-diamidino-2-phenylindole (DAPI). No vimentin staining was observed in primary epithelial cell cultures (data not shown).
Figure Legend Snippet: Characterization of human primary prostate epithelial cell cultures. Notes: ( A ) Primary outgrowth of epithelial cells derived from a biopsy specimen. ( B ) Primary epithelial cell colony. ( C ) Characterization of epithelial cells: green fluorescence shows high-molecular-weight cytokeratins; blue fluorescence shows nuclei reactive with 4′,6-diamidino-2-phenylindole (DAPI). No vimentin staining was observed in primary epithelial cell cultures (data not shown).

Techniques Used: Derivative Assay, Fluorescence, Molecular Weight, Staining

5) Product Images from "Elevated PDK1 Expression Drives PI3K/AKT/MTOR Signaling Promotes Radiation-Resistant and Dedifferentiated Phenotype of Hepatocellular Carcinoma"

Article Title: Elevated PDK1 Expression Drives PI3K/AKT/MTOR Signaling Promotes Radiation-Resistant and Dedifferentiated Phenotype of Hepatocellular Carcinoma

Journal: Cells

doi: 10.3390/cells9030746

PDK1-dependent radioresistance is associated with the enhanced metastatic and cancer stem cell-like phenotypes of HCC cells. Representative photo-images (upper panel) and histograms (lower panel) comparing ( A ) migration and ( B ) invasion potential between Mahlavu and Mahlavu-R cells. Scale bar: 100 μm, 10× objective. ( C ) Representative photo-images (left panel) comparing clonogenicity between Huh7-R, Mahlavu-R, Hep3B-R and their wild-type counterparts. Scale bar: 100 μm, 10× objective. ( D ) Representative Western blot images of the differential expression of p-PDK1, PDK1, E-cadherin, N -cadherin, Vimentin or Snail protein in wild-type and IR-resistant Hep3B, Mahlavu or Huh7 cells. β-actin was used as a loading control. ( E ) Representative photo-images (upper panel) and histograms (lower panel) comparing the tumorsphere formation potential between wild-type (WT) and IR-resistant Huh7 or Mahlavu cells. Scale bar: 100 μm, 10× objective. ( F ) Representative immunofluorescence staining images of SOX2 and OCT4A expression in WT and IR-resistant Huh7 or Mahlavu cells. DAPI served as nuclear marker. Scale bar: 100 μm, 10× objective. * p
Figure Legend Snippet: PDK1-dependent radioresistance is associated with the enhanced metastatic and cancer stem cell-like phenotypes of HCC cells. Representative photo-images (upper panel) and histograms (lower panel) comparing ( A ) migration and ( B ) invasion potential between Mahlavu and Mahlavu-R cells. Scale bar: 100 μm, 10× objective. ( C ) Representative photo-images (left panel) comparing clonogenicity between Huh7-R, Mahlavu-R, Hep3B-R and their wild-type counterparts. Scale bar: 100 μm, 10× objective. ( D ) Representative Western blot images of the differential expression of p-PDK1, PDK1, E-cadherin, N -cadherin, Vimentin or Snail protein in wild-type and IR-resistant Hep3B, Mahlavu or Huh7 cells. β-actin was used as a loading control. ( E ) Representative photo-images (upper panel) and histograms (lower panel) comparing the tumorsphere formation potential between wild-type (WT) and IR-resistant Huh7 or Mahlavu cells. Scale bar: 100 μm, 10× objective. ( F ) Representative immunofluorescence staining images of SOX2 and OCT4A expression in WT and IR-resistant Huh7 or Mahlavu cells. DAPI served as nuclear marker. Scale bar: 100 μm, 10× objective. * p

Techniques Used: Migration, Western Blot, Expressing, Immunofluorescence, Staining, Marker

6) Product Images from "Iron Exposure and the Cellular Mechanisms Linked to Neuron Degeneration in Adult Mice"

Article Title: Iron Exposure and the Cellular Mechanisms Linked to Neuron Degeneration in Adult Mice

Journal: Cells

doi: 10.3390/cells8020198

( A ) Immunofluorescence of Aβ and NeuN. Scale bar = 25 μm, ( A1 , A2 ) Quantitative analyses of Neun-positive staining and Aβ-positive staining. ( B ) Nissl staining in the mouse brain. Scale bar = 25 μm. ( B1 ) Quantitative analyses of Nissl-positive staining, n = 8. * p
Figure Legend Snippet: ( A ) Immunofluorescence of Aβ and NeuN. Scale bar = 25 μm, ( A1 , A2 ) Quantitative analyses of Neun-positive staining and Aβ-positive staining. ( B ) Nissl staining in the mouse brain. Scale bar = 25 μm. ( B1 ) Quantitative analyses of Nissl-positive staining, n = 8. * p

Techniques Used: Immunofluorescence, Staining

7) Product Images from "Role of mucosa in generating spontaneous activity in the guinea pig seminal vesicle"

Article Title: Role of mucosa in generating spontaneous activity in the guinea pig seminal vesicle

Journal: The Journal of Physiology

doi: 10.1113/JP273872

Distribution of the interstitial cells in the subepithelium of SV mucosa preparations A , an illustration of dissected SV mucosa from the muscular layer used for the whole mount preparation in B , C and F . Serial images were obtained from the basal (subepithelial) side of the mucosa preparations via confocal microscopy. B , serial images of a whole mount SV mucosa preparation immunolabelled with anti‐pancytokeratin (red: a marker of epithelial cell) and vimentin (green: a marker of interstitial cell). ‘a + 5 μm’ in the right image indicates that the image was obtained 5 μm from the basal side of ‘a’. Vimentin‐immunoreactive (IR) cells located beneath the epithelial layer. C , double immunolabelling with anti‐c‐Kit and α‐SMA antibodies in another whole mount SV mucosa preparation. α‐SMA‐IR (red) cells were not found except for the blood vessels, and a few oval‐shaped c‐Kit‐IR (green) cells are observed. Double immunolabelled cross‐sections of whole tissue of SV ( D ) and muscular layer of gastric antrum ( E ) produced under the same protocol. In SV wall, non‐significant signals of c‐Kit (green) were observed, while interstitial cells of Cajal immunolabelled with c‐Kit antibody were observed in the stomach. LP, lamina propria; M, musclular layer. F , serial images of a whole mount SV mucosa preparation. PDGFRα‐IR (green) cells are found beneath the epithelial layer (a + 4.2 μm, a + 6.2 μm). Scale bars = 40 μm.
Figure Legend Snippet: Distribution of the interstitial cells in the subepithelium of SV mucosa preparations A , an illustration of dissected SV mucosa from the muscular layer used for the whole mount preparation in B , C and F . Serial images were obtained from the basal (subepithelial) side of the mucosa preparations via confocal microscopy. B , serial images of a whole mount SV mucosa preparation immunolabelled with anti‐pancytokeratin (red: a marker of epithelial cell) and vimentin (green: a marker of interstitial cell). ‘a + 5 μm’ in the right image indicates that the image was obtained 5 μm from the basal side of ‘a’. Vimentin‐immunoreactive (IR) cells located beneath the epithelial layer. C , double immunolabelling with anti‐c‐Kit and α‐SMA antibodies in another whole mount SV mucosa preparation. α‐SMA‐IR (red) cells were not found except for the blood vessels, and a few oval‐shaped c‐Kit‐IR (green) cells are observed. Double immunolabelled cross‐sections of whole tissue of SV ( D ) and muscular layer of gastric antrum ( E ) produced under the same protocol. In SV wall, non‐significant signals of c‐Kit (green) were observed, while interstitial cells of Cajal immunolabelled with c‐Kit antibody were observed in the stomach. LP, lamina propria; M, musclular layer. F , serial images of a whole mount SV mucosa preparation. PDGFRα‐IR (green) cells are found beneath the epithelial layer (a + 4.2 μm, a + 6.2 μm). Scale bars = 40 μm.

Techniques Used: Confocal Microscopy, Marker, Produced

8) Product Images from "CDX2 Stimulates the Proliferation of Porcine Intestinal Epithelial Cells by Activating the mTORC1 and Wnt/β-Catenin Signaling Pathways"

Article Title: CDX2 Stimulates the Proliferation of Porcine Intestinal Epithelial Cells by Activating the mTORC1 and Wnt/β-Catenin Signaling Pathways

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms18112447

CDX2 knockdown inhibited both the mTORC1 and Wnt/β-catenin pathways. ( a,b ) Western blot analysis of the mTORC1 pathway activity after CDX2 knockdown in IPEC-J2 cells with quantification ( n = 3); ( c,d ) western blot of Wnt/β-catenin pathway related proteins after CDX2 knockdown with quantification ( n = 3). AU: arbitrary unit. Negative Control: negative control group; Knockdown: CDX2-siRNA-002 group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM ( n = 3); * p
Figure Legend Snippet: CDX2 knockdown inhibited both the mTORC1 and Wnt/β-catenin pathways. ( a,b ) Western blot analysis of the mTORC1 pathway activity after CDX2 knockdown in IPEC-J2 cells with quantification ( n = 3); ( c,d ) western blot of Wnt/β-catenin pathway related proteins after CDX2 knockdown with quantification ( n = 3). AU: arbitrary unit. Negative Control: negative control group; Knockdown: CDX2-siRNA-002 group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM ( n = 3); * p

Techniques Used: Western Blot, Activity Assay, Negative Control

Caudal type homeobox 2 (CDX2) overexpressed in IPEC-J2 cells and increased cell proliferation. ( a ) Representative immunofluorescence images of control and overexpression groups 48 h after seeding, labeled with DAPI (blue) and CDX2 antibody (red) (200×). Scale bar: 100 μm. ( b,c ) CDX2 mRNA abundance ( n = 6) and protein expression ( n = 3) in the control and overexpression group. AU: arbitrary unit; ( d,e ) the OD value and cell number were assessed by MTT assay ( n = 20) and cell counting ( n = 6), respectively. Control: control group; Overexpression: CDX2 overexpression group. Representative results of three independent experiments are shown as the mean ± SEM; * p
Figure Legend Snippet: Caudal type homeobox 2 (CDX2) overexpressed in IPEC-J2 cells and increased cell proliferation. ( a ) Representative immunofluorescence images of control and overexpression groups 48 h after seeding, labeled with DAPI (blue) and CDX2 antibody (red) (200×). Scale bar: 100 μm. ( b,c ) CDX2 mRNA abundance ( n = 6) and protein expression ( n = 3) in the control and overexpression group. AU: arbitrary unit; ( d,e ) the OD value and cell number were assessed by MTT assay ( n = 20) and cell counting ( n = 6), respectively. Control: control group; Overexpression: CDX2 overexpression group. Representative results of three independent experiments are shown as the mean ± SEM; * p

Techniques Used: Immunofluorescence, Over Expression, Labeling, Expressing, MTT Assay, Cell Counting

In IPEC-J2 cells, CDX2 promotes cell proliferation via activating the mTORC1 and Wnt/β-catenin pathways; specific antagonists of the mTORC1 and Wnt/β-catenin pathways, namely rapamycin or XAV939 respectively, decrease cell proliferation and inhibit both pathways simultaneously.
Figure Legend Snippet: In IPEC-J2 cells, CDX2 promotes cell proliferation via activating the mTORC1 and Wnt/β-catenin pathways; specific antagonists of the mTORC1 and Wnt/β-catenin pathways, namely rapamycin or XAV939 respectively, decrease cell proliferation and inhibit both pathways simultaneously.

Techniques Used:

Specific antagonists inhibited both the mTORC1 and Wnt/β-catenin pathways in CDX2 overexpressed IPEC-J2 cells. ( a – d ) Western blot analysis of the mTORC1 and Wnt/β-catenin pathways activity after treated with rapamycin in CDX2 overexpressed IPEC-J2 cells with quantification ( n = 3); ( e – h ) western blot of the mTORC1 and Wnt/β-catenin pathways related proteins after treated with XAV939 in CDX2 overexpressed IPEC-J2 cells with quantification ( n = 3); ( i – l ) protein levels of the mTORC1 and Wnt/β-catenin pathways were measured by Western blot after treated with rapamycin and XAV939 in combination with quantification ( n = 3). AU: arbitrary unit. As rapamycin and XAV939 were dissolved in DMSO, the DMSO-treated cell strain was the control group. DMSO: DMSO treatment group; Rap.: rapamycin treatment group; XAV.: XAV939 treatment group; Rap. + XAV.: combination of rapamycin and XAV939 treatment group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM ( n = 3); * p
Figure Legend Snippet: Specific antagonists inhibited both the mTORC1 and Wnt/β-catenin pathways in CDX2 overexpressed IPEC-J2 cells. ( a – d ) Western blot analysis of the mTORC1 and Wnt/β-catenin pathways activity after treated with rapamycin in CDX2 overexpressed IPEC-J2 cells with quantification ( n = 3); ( e – h ) western blot of the mTORC1 and Wnt/β-catenin pathways related proteins after treated with XAV939 in CDX2 overexpressed IPEC-J2 cells with quantification ( n = 3); ( i – l ) protein levels of the mTORC1 and Wnt/β-catenin pathways were measured by Western blot after treated with rapamycin and XAV939 in combination with quantification ( n = 3). AU: arbitrary unit. As rapamycin and XAV939 were dissolved in DMSO, the DMSO-treated cell strain was the control group. DMSO: DMSO treatment group; Rap.: rapamycin treatment group; XAV.: XAV939 treatment group; Rap. + XAV.: combination of rapamycin and XAV939 treatment group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM ( n = 3); * p

Techniques Used: Western Blot, Activity Assay

Specific antagonists decreased cell proliferation and the protein level of CDX2 in CDX2 overexpressed IPEC-J2 cells. ( a,b ) OD values and cell numbers were assessed by MTT assay ( n = 20) and cell counting ( n = 6), respectively; ( c – e ) protein levels of CDX2 were measured by Western blot after treated with rapamycin and XAV939 alone or in combination with quantification ( n = 3). AU: arbitrary unit. DMSO: DMSO treatment group; Rap.: rapamycin treatment group; XAV.: XAV939 treatment group; Rap. + XAV.: combination of rapamycin and XAV939 treatment group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM; bars without the same letter indicate a significant difference; * p
Figure Legend Snippet: Specific antagonists decreased cell proliferation and the protein level of CDX2 in CDX2 overexpressed IPEC-J2 cells. ( a,b ) OD values and cell numbers were assessed by MTT assay ( n = 20) and cell counting ( n = 6), respectively; ( c – e ) protein levels of CDX2 were measured by Western blot after treated with rapamycin and XAV939 alone or in combination with quantification ( n = 3). AU: arbitrary unit. DMSO: DMSO treatment group; Rap.: rapamycin treatment group; XAV.: XAV939 treatment group; Rap. + XAV.: combination of rapamycin and XAV939 treatment group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM; bars without the same letter indicate a significant difference; * p

Techniques Used: MTT Assay, Cell Counting, Western Blot

CDX2 overexpression activated both the mTORC1 and Wnt/β-catenin pathways. ( a,b ) Western blot analysis of the mTORC1 pathway activity after CDX2 overexpressed in IPEC-J2 cells with quantification ( n = 3); ( c,d ) western blot of Wnt/β-catenin pathway related proteins after CDX2 overexpressed with quantification ( n = 3). AU: arbitrary unit. Control: control group; Overexpression: CDX2 overexpression group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM; * p
Figure Legend Snippet: CDX2 overexpression activated both the mTORC1 and Wnt/β-catenin pathways. ( a,b ) Western blot analysis of the mTORC1 pathway activity after CDX2 overexpressed in IPEC-J2 cells with quantification ( n = 3); ( c,d ) western blot of Wnt/β-catenin pathway related proteins after CDX2 overexpressed with quantification ( n = 3). AU: arbitrary unit. Control: control group; Overexpression: CDX2 overexpression group. Representative results of three independent experiments are shown. Data are expressed as the mean ± SEM; * p

Techniques Used: Over Expression, Western Blot, Activity Assay

CDX2 knockdown in IPEC-J2 cells reduced cell proliferation. ( a ) The effect of three siRNAs on CDX2 mRNA abundance was measured by real-time PCR 48 h post-transfection. Blank: control group; NC: negative control group; siRNA-001: CDX2-siRNA-001 group; siRNA-002: CDX2-siRNA-002 group; siRNA-003: CDX2-siRNA-003 group; ( b ) the effect of siRNA-002 transfection time on CDX2 mRNA abundance was measured by real-time PCR. Data are expressed as the mean ± SEM ( n = 6). The bars without same letters indicate a significant difference ( p
Figure Legend Snippet: CDX2 knockdown in IPEC-J2 cells reduced cell proliferation. ( a ) The effect of three siRNAs on CDX2 mRNA abundance was measured by real-time PCR 48 h post-transfection. Blank: control group; NC: negative control group; siRNA-001: CDX2-siRNA-001 group; siRNA-002: CDX2-siRNA-002 group; siRNA-003: CDX2-siRNA-003 group; ( b ) the effect of siRNA-002 transfection time on CDX2 mRNA abundance was measured by real-time PCR. Data are expressed as the mean ± SEM ( n = 6). The bars without same letters indicate a significant difference ( p

Techniques Used: Real-time Polymerase Chain Reaction, Transfection, Negative Control

9) Product Images from "Novel elucidation and treatment of pancreatic chronic graft-versus-host disease in mice"

Article Title: Novel elucidation and treatment of pancreatic chronic graft-versus-host disease in mice

Journal: Royal Society Open Science

doi: 10.1098/rsos.181067

EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in cGVHD-affected pancreas and its syngeneic control counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane1: syngeneic control subject, lane 2: cGVHD-affected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S9. ( c ) The corresponding quantitative analysis of the protein bands was conducted. cGVHD-affected pancreas (red) and its syngeneic control counterpart (blue). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., control: n = 5, cGVHD: n = 5.
Figure Legend Snippet: EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in cGVHD-affected pancreas and its syngeneic control counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane1: syngeneic control subject, lane 2: cGVHD-affected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S9. ( c ) The corresponding quantitative analysis of the protein bands was conducted. cGVHD-affected pancreas (red) and its syngeneic control counterpart (blue). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., control: n = 5, cGVHD: n = 5.

Techniques Used: Immunostaining, Staining, Fluorescence

EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in the PBA-treated pancreas and its vehicle-treated counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane 1: PBA-injected pancreas, lane 2: vehicle-injected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S19. ( c ) The corresponding quantitative analysis of the protein bands was conducted. PBA-medicated pancreas (blue) and its vehicle-medicated counterpart (red). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., PBA: n = 6, vehicle: n = 6.
Figure Legend Snippet: EMT markers in the cGVHD-affected pancreas. ( a ) Immunostaining for E-cadherin in the PBA-treated pancreas and its vehicle-treated counterpart. E-cadherin and cell nuclei are stained green and blue, respectively. The fluorescence images were at 200× magnification, and the scale bar is 20 µm. ( b ) Immunoblot analysis of E-cadherin and α-SMA was carried out (lane 1: PBA-injected pancreas, lane 2: vehicle-injected pancreas). Cropped blots are displayed, and the corresponding full-length gels are shown in electronic supplementary material, figure S19. ( c ) The corresponding quantitative analysis of the protein bands was conducted. PBA-medicated pancreas (blue) and its vehicle-medicated counterpart (red). Results are representative of two independently performed experiments with similar results. The data are presented as means ± s.d., PBA: n = 6, vehicle: n = 6.

Techniques Used: Immunostaining, Staining, Fluorescence, Injection

10) Product Images from "Cytotoxic Metabolites from Callyspongia siphonella Display Antiproliferative Activity by Inducing Apoptosis in HCT-116 Cells"

Article Title: Cytotoxic Metabolites from Callyspongia siphonella Display Antiproliferative Activity by Inducing Apoptosis in HCT-116 Cells

Journal: Pharmacognosy Magazine

doi: 10.4103/0973-1296.203970

Effect of sipholenol-A and sipholenol-L at their respective IC 50 concentration on the levels of cell cycle regulatory proteins in HCT-116 cells after 24 h of incubation. Cyclin-B1, cyclin-D1, and cleaved caspase-3 were assessed inmmunocytochemically (red), and cells were counterstained with nuclear DAPI stain (blue)
Figure Legend Snippet: Effect of sipholenol-A and sipholenol-L at their respective IC 50 concentration on the levels of cell cycle regulatory proteins in HCT-116 cells after 24 h of incubation. Cyclin-B1, cyclin-D1, and cleaved caspase-3 were assessed inmmunocytochemically (red), and cells were counterstained with nuclear DAPI stain (blue)

Techniques Used: Concentration Assay, Incubation, Staining

Quantitation of fluorescence intensity sipholenol-A and sipholenol-L on cylin-B1, cyclin-D1, and cleaved caspase-3. AU: arbitrary units. *denotes significantly different from control at P
Figure Legend Snippet: Quantitation of fluorescence intensity sipholenol-A and sipholenol-L on cylin-B1, cyclin-D1, and cleaved caspase-3. AU: arbitrary units. *denotes significantly different from control at P

Techniques Used: Quantitation Assay, Fluorescence

11) Product Images from "Comparative characteristics of laryngeal-resident mesenchymal stromal cell populations isolated from distinct sites in the rat larynx"

Article Title: Comparative characteristics of laryngeal-resident mesenchymal stromal cell populations isolated from distinct sites in the rat larynx

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-017-0650-y

Isolation and validation of laryngeal tissue-resident clonal cells. a Clonal cells purified from laryngeal subsites of EM, LP and MF visualized by phase-contrast microscopy. Scale bars, 100 and 20 μm. Representative immunofluorescence staining images of laryngeal clonal cells expressing mesenchymal markers vimentin and α-SMA. b Expression of the genes Krt19 (epithelial cell marker) and Vwf (endothelial cell marker) was not detected, whereas that of Vim and Acta2 was detected in all laryngeal clonal cells. c MF stellate cells were validated to contain lipid droplets (arrow) by phase-contrast microscopy. Scale bars, 25 μm. These cells demonstrated vitamin A (retinoid) autofluorescence as assessed by d fluorescence microscopy (scale bars, 10 μm) and e retinoid-based FACS sorting. EM epiglottic mucosa, LP lamina propria, MF macula flava, α-SMA α-smooth muscle actin, FSC forward scatter
Figure Legend Snippet: Isolation and validation of laryngeal tissue-resident clonal cells. a Clonal cells purified from laryngeal subsites of EM, LP and MF visualized by phase-contrast microscopy. Scale bars, 100 and 20 μm. Representative immunofluorescence staining images of laryngeal clonal cells expressing mesenchymal markers vimentin and α-SMA. b Expression of the genes Krt19 (epithelial cell marker) and Vwf (endothelial cell marker) was not detected, whereas that of Vim and Acta2 was detected in all laryngeal clonal cells. c MF stellate cells were validated to contain lipid droplets (arrow) by phase-contrast microscopy. Scale bars, 25 μm. These cells demonstrated vitamin A (retinoid) autofluorescence as assessed by d fluorescence microscopy (scale bars, 10 μm) and e retinoid-based FACS sorting. EM epiglottic mucosa, LP lamina propria, MF macula flava, α-SMA α-smooth muscle actin, FSC forward scatter

Techniques Used: Isolation, Purification, Microscopy, Immunofluorescence, Staining, Expressing, Marker, Fluorescence, FACS

12) Product Images from "Autophagy Is Impaired in Neutrophils from Streptozotocin-Induced Diabetic Rats"

Article Title: Autophagy Is Impaired in Neutrophils from Streptozotocin-Induced Diabetic Rats

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00024

Contents of LC3BI and LC3BII in neutrophils before and after PMA stimulus . Neutrophils from diabetic rats had low cleavage of LC3BI to LC3BII when compared to the control group (A) . Graph (B) presents mean OD ± SEM of the bands from six animals per group.
Figure Legend Snippet: Contents of LC3BI and LC3BII in neutrophils before and after PMA stimulus . Neutrophils from diabetic rats had low cleavage of LC3BI to LC3BII when compared to the control group (A) . Graph (B) presents mean OD ± SEM of the bands from six animals per group.

Techniques Used:

13) Product Images from "The Absence of Indoleamine 2,3-Dioxygenase Inhibits Retinal Capillary Degeneration in Diabetic Mice"

Article Title: The Absence of Indoleamine 2,3-Dioxygenase Inhibits Retinal Capillary Degeneration in Diabetic Mice

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.17-22702

Characterization of isolated HREC. Isolated HREC were characterized by (A) PECAM-1 immunostaining and (B) an assay of DiL-Ac-LDL uptake. The images are representative of three independent experiments. Scale bar: 100 μm.
Figure Legend Snippet: Characterization of isolated HREC. Isolated HREC were characterized by (A) PECAM-1 immunostaining and (B) an assay of DiL-Ac-LDL uptake. The images are representative of three independent experiments. Scale bar: 100 μm.

Techniques Used: Isolation, Immunostaining

14) Product Images from "Iron Exposure and the Cellular Mechanisms Linked to Neuron Degeneration in Adult Mice"

Article Title: Iron Exposure and the Cellular Mechanisms Linked to Neuron Degeneration in Adult Mice

Journal: Cells

doi: 10.3390/cells8020198

( A ) Immunofluorescence of Aβ and NeuN. Scale bar = 25 μm, ( A1 , A2 ) Quantitative analyses of Neun-positive staining and Aβ-positive staining. ( B ) Nissl staining in the mouse brain. Scale bar = 25 μm. ( B1 ) Quantitative analyses of Nissl-positive staining, n = 8. * p
Figure Legend Snippet: ( A ) Immunofluorescence of Aβ and NeuN. Scale bar = 25 μm, ( A1 , A2 ) Quantitative analyses of Neun-positive staining and Aβ-positive staining. ( B ) Nissl staining in the mouse brain. Scale bar = 25 μm. ( B1 ) Quantitative analyses of Nissl-positive staining, n = 8. * p

Techniques Used: Immunofluorescence, Staining

15) Product Images from "Overexpression of Mitochondria Mediator Gene TRIAP1 by miR-320b Loss Is Associated with Progression in Nasopharyngeal Carcinoma"

Article Title: Overexpression of Mitochondria Mediator Gene TRIAP1 by miR-320b Loss Is Associated with Progression in Nasopharyngeal Carcinoma

Journal: PLoS Genetics

doi: 10.1371/journal.pgen.1006183

TRIAP1 mediates the effects of miR-320b on NPC cell proliferation, mitochondrial fragmentation and apoptosis. ( A-F ) CNE-2 and SUNE-1 cells were co-transfected with a miR-320b mimic or miR-Ctrl and either the empty vector (Vector) or plasmid overexpressing TRIAP1. ( A ) MTT assay showing that recovery of TRIAP1 partially rescues the inhibitory effects of miR-320b on cell proliferation. ( B-F ) Flow cytometric analysis ( B-C ), caspase-3/7 ( D ) and immunofluorescent staining ( E-F ) assay showing that restoration of TRIAP1 reverses the promoting effects of miR-320b on mitochondrial fragmentation, apoptosis and cytochrome c release from mitochondria. Scale bar, 10 μm. Each experiment was independently repeated at least three times. The data are presented as the mean ± s.d. Student’s t -test, * P
Figure Legend Snippet: TRIAP1 mediates the effects of miR-320b on NPC cell proliferation, mitochondrial fragmentation and apoptosis. ( A-F ) CNE-2 and SUNE-1 cells were co-transfected with a miR-320b mimic or miR-Ctrl and either the empty vector (Vector) or plasmid overexpressing TRIAP1. ( A ) MTT assay showing that recovery of TRIAP1 partially rescues the inhibitory effects of miR-320b on cell proliferation. ( B-F ) Flow cytometric analysis ( B-C ), caspase-3/7 ( D ) and immunofluorescent staining ( E-F ) assay showing that restoration of TRIAP1 reverses the promoting effects of miR-320b on mitochondrial fragmentation, apoptosis and cytochrome c release from mitochondria. Scale bar, 10 μm. Each experiment was independently repeated at least three times. The data are presented as the mean ± s.d. Student’s t -test, * P

Techniques Used: Transfection, Plasmid Preparation, MTT Assay, Flow Cytometry, Staining

TRIAP1 regulates apoptosis through controlling the release of cytochrome c . A and B , Release of cytochrome c is increased upon TRIAP1 knockdown. Representative images of mitochondria, cytochrome c ( A ) and TRIAP1 ( B ) subcellular locations for CNE-2 (left panel) and SUNE-1 (right panel) cells transiently transfected with siSCR, siTRIAP1-1 or siTRIAP1-2 after staining with MitoTracker Red, cytochrome c or TRIAP1 primary antibodies. Scale bar, 10 μm. ( C ) Caspase-3/7 assay for cells with TRIAP1 knockdown. Each experiment was independently repeated at least three times. The data are presented as the mean ± s.d. Student’s t -test, * P
Figure Legend Snippet: TRIAP1 regulates apoptosis through controlling the release of cytochrome c . A and B , Release of cytochrome c is increased upon TRIAP1 knockdown. Representative images of mitochondria, cytochrome c ( A ) and TRIAP1 ( B ) subcellular locations for CNE-2 (left panel) and SUNE-1 (right panel) cells transiently transfected with siSCR, siTRIAP1-1 or siTRIAP1-2 after staining with MitoTracker Red, cytochrome c or TRIAP1 primary antibodies. Scale bar, 10 μm. ( C ) Caspase-3/7 assay for cells with TRIAP1 knockdown. Each experiment was independently repeated at least three times. The data are presented as the mean ± s.d. Student’s t -test, * P

Techniques Used: Transfection, Staining

16) Product Images from "Proteomic analysis of energy metabolism and signal transduction in irradiated melanoma cells"

Article Title: Proteomic analysis of energy metabolism and signal transduction in irradiated melanoma cells

Journal: International Journal of Ophthalmology

doi: 10.3980/j.issn.2222-3959.2013.03.06

Post-irradiation cell phenotypes A: FACS cell cycle analysis; B: Lactate production at 15-hour post-irradiation; C: Ki67 immunofluorescent staining quantitation (left) and representative images (right).
Figure Legend Snippet: Post-irradiation cell phenotypes A: FACS cell cycle analysis; B: Lactate production at 15-hour post-irradiation; C: Ki67 immunofluorescent staining quantitation (left) and representative images (right).

Techniques Used: Irradiation, FACS, Cell Cycle Assay, Staining, Quantitation Assay

17) Product Images from "Primary cilia modulate balance of canonical and non-canonical Wnt signaling responses in the injured kidney"

Article Title: Primary cilia modulate balance of canonical and non-canonical Wnt signaling responses in the injured kidney

Journal: Fibrogenesis & Tissue Repair

doi: 10.1186/s13069-015-0024-y

Stunted cilia and canonical Wnt signaling in experimental obstructive nephropathy. ( A ) Paraffin-embedded kidney sections were labeled with antibodies to acetylated α-tubulin (Ac-α-tubulin) (green). Nuclei were labeled with DAPI (blue). Staining was analyzed using a confocal microscope. The pictures display representative confocal photomicrographs of labeled sections of sham-operated mice (left) and of kidneys 3 days (middle) and 7 days (right) after ureter obstruction. Scale bars: 10 μm. Arrows indicate normal cilia and arrowheads indicate stunted cilia. ( B ) The graph summarizes the relative distribution of normal and stunted cilia in each group. ( C ) Immunofluorescence double-labeling experiments were performed using specific antibodies to acetylated α-tubulin (cilia) and non-phosphorylated β-catenin (active canonical Wnt signaling). Nuclei were stained with DAPI. The pictures display representative photomicrographs of sections from sham-operated control mice (left) and post-obstructive (days 3 and 7) kidneys. Non-phosphorylated β-catenin immunostaining was most abundant in tubules with stunted cilia. Arrows indicate normal cilia and arrowheads display stunted cilia. Scale bars: 10 μm. ( D ) The graph shows relative distribution of normal cilia and stunted cilia regarding to β-catenin expression in each group. ( E ) Non-fibrotic and fibrotic kidney sections from human biopsies were labeled with acetylated α-tubulin (cilia) and non-phosphorylated β-catenin. Pictures display representative confocal photomicrographs of each sample. Arrows indicate normal cilia and arrowheads indicate stunted cilia. Scale bars: 10 μm. UUO, unilateral ureteral obstruction; DAPI, 4′,6-diamidino-2-phenylindole.
Figure Legend Snippet: Stunted cilia and canonical Wnt signaling in experimental obstructive nephropathy. ( A ) Paraffin-embedded kidney sections were labeled with antibodies to acetylated α-tubulin (Ac-α-tubulin) (green). Nuclei were labeled with DAPI (blue). Staining was analyzed using a confocal microscope. The pictures display representative confocal photomicrographs of labeled sections of sham-operated mice (left) and of kidneys 3 days (middle) and 7 days (right) after ureter obstruction. Scale bars: 10 μm. Arrows indicate normal cilia and arrowheads indicate stunted cilia. ( B ) The graph summarizes the relative distribution of normal and stunted cilia in each group. ( C ) Immunofluorescence double-labeling experiments were performed using specific antibodies to acetylated α-tubulin (cilia) and non-phosphorylated β-catenin (active canonical Wnt signaling). Nuclei were stained with DAPI. The pictures display representative photomicrographs of sections from sham-operated control mice (left) and post-obstructive (days 3 and 7) kidneys. Non-phosphorylated β-catenin immunostaining was most abundant in tubules with stunted cilia. Arrows indicate normal cilia and arrowheads display stunted cilia. Scale bars: 10 μm. ( D ) The graph shows relative distribution of normal cilia and stunted cilia regarding to β-catenin expression in each group. ( E ) Non-fibrotic and fibrotic kidney sections from human biopsies were labeled with acetylated α-tubulin (cilia) and non-phosphorylated β-catenin. Pictures display representative confocal photomicrographs of each sample. Arrows indicate normal cilia and arrowheads indicate stunted cilia. Scale bars: 10 μm. UUO, unilateral ureteral obstruction; DAPI, 4′,6-diamidino-2-phenylindole.

Techniques Used: Labeling, Staining, Microscopy, Mouse Assay, Immunofluorescence, Immunostaining, Expressing

Renal expression of Wnt ligand genes upon unilateral ureter obstruction ( A-K ). We isolated total RNA from whole kidney tissues 3 and 7 days after ureter ligation and analyzed expression of putative Wnt ligand genes by quantitative real-time PCR. The bar graphs display relative mRNA expression of indicated Wnt ligand genes upon ureter ligation in relation to kidneys of sham-operated control mice. The arrows highlight robust expression of β-catenin in dilated tubules. Data is presented as means ± SD. * P
Figure Legend Snippet: Renal expression of Wnt ligand genes upon unilateral ureter obstruction ( A-K ). We isolated total RNA from whole kidney tissues 3 and 7 days after ureter ligation and analyzed expression of putative Wnt ligand genes by quantitative real-time PCR. The bar graphs display relative mRNA expression of indicated Wnt ligand genes upon ureter ligation in relation to kidneys of sham-operated control mice. The arrows highlight robust expression of β-catenin in dilated tubules. Data is presented as means ± SD. * P

Techniques Used: Expressing, Isolation, Ligation, Real-time Polymerase Chain Reaction, Mouse Assay

Reciprocal responses against Wnt in ciliated and non-ciliated cells. We compared induction of canonical and non-canonical Wnt target genes in response to Wnt3a in cultured ciliated and non-ciliated HK2 tubular epithelial cells. The graphs display relative mRNA expression of indicated genes in ciliated versus non-ciliated cells with or without Wnt3a in culture media. ( A ) Ciliated and non-ciliated HK2 cells were methanol fixed and were labeled with antibodies against acetylated α-tubulin (cilia) and non-phosphorylated β-catenin. Higher expression of active β-catenin was observed in Wnt3a-treated non-ciliated cells. Arrows highlight normal cilia on epithelial cell. Scale bars: 10 μm. ( B - E ) Expression of canonical Wnt signaling target genes (AXIN2, WISP2, TP53, CCND1) was substantially induced in ciliated cells but not in non-ciliated cells. ( F - I ) Addition of Wnt3a to culture media induced expression of non-canonical target genes in ciliated cells but not in non-ciliated HK2 cells (CAMK2A, PLCB1, DAAM1, PTK7). Data is presented as means ± SD. * P
Figure Legend Snippet: Reciprocal responses against Wnt in ciliated and non-ciliated cells. We compared induction of canonical and non-canonical Wnt target genes in response to Wnt3a in cultured ciliated and non-ciliated HK2 tubular epithelial cells. The graphs display relative mRNA expression of indicated genes in ciliated versus non-ciliated cells with or without Wnt3a in culture media. ( A ) Ciliated and non-ciliated HK2 cells were methanol fixed and were labeled with antibodies against acetylated α-tubulin (cilia) and non-phosphorylated β-catenin. Higher expression of active β-catenin was observed in Wnt3a-treated non-ciliated cells. Arrows highlight normal cilia on epithelial cell. Scale bars: 10 μm. ( B - E ) Expression of canonical Wnt signaling target genes (AXIN2, WISP2, TP53, CCND1) was substantially induced in ciliated cells but not in non-ciliated cells. ( F - I ) Addition of Wnt3a to culture media induced expression of non-canonical target genes in ciliated cells but not in non-ciliated HK2 cells (CAMK2A, PLCB1, DAAM1, PTK7). Data is presented as means ± SD. * P

Techniques Used: Cell Culture, Expressing, Labeling

β-catenin immunolocalization in experimental obstructive nephropathy. We labeled kidneys of sham-operated mice and of mice which had been challenged with UUO with antibodies specific to ( A ) total β-catenin or with antibodies specific for ( B ) non-phosphorylated β-catenin (indicative of active canonical signaling). The pictures display representative photomicrographs of each groups, positive immunostaining is indicated by brown precipitate. The arrows highlight robust expression of β-catenin in dilated tubules. Scale bars: 100 μm. UUO, unilateral ureteral obstruction.
Figure Legend Snippet: β-catenin immunolocalization in experimental obstructive nephropathy. We labeled kidneys of sham-operated mice and of mice which had been challenged with UUO with antibodies specific to ( A ) total β-catenin or with antibodies specific for ( B ) non-phosphorylated β-catenin (indicative of active canonical signaling). The pictures display representative photomicrographs of each groups, positive immunostaining is indicated by brown precipitate. The arrows highlight robust expression of β-catenin in dilated tubules. Scale bars: 100 μm. UUO, unilateral ureteral obstruction.

Techniques Used: Labeling, Mouse Assay, Immunostaining, Expressing

18) Product Images from "Age-related macular degeneration-like phenotypic features develop at the early ages of Cxcr5/Nrf2 double knockout mice: An accelerated AMD model"

Article Title: Age-related macular degeneration-like phenotypic features develop at the early ages of Cxcr5/Nrf2 double knockout mice: An accelerated AMD model

Journal: bioRxiv

doi: 10.1101/868851

Accelerated retinal degeneration and cell apoptosis in the adult CXCR5 -/- . NRF2 -/- mice. The 6-month-old C5BL6 WT mice, CXCR5 -/- KO mice, and CXCR5 -/- .NRF2 -/- DKO mice retinal flatmounts were used for staining and quantification. (A-D) Retinal flatmounts stained with peanut agglutinin (PNA) lectin (A), MAP2 (B), cleaved caspase 3 at the ganglion cell layer (C) and the photoreceptor cell layer depth (D). The quantitative results of PNA lectin (+) photoreceptors (E), MAP2 (+) retinal ganglion cells (F). Caspase 3 (+) ganglion cells layer (G), and Caspase 3 (+) photoreceptor layer depths (H). The values of cell numbers per 100 µm 2 were averaged from 4 retinal samples (n = 4). P values were denoted: n.s. P > 0.05; * P
Figure Legend Snippet: Accelerated retinal degeneration and cell apoptosis in the adult CXCR5 -/- . NRF2 -/- mice. The 6-month-old C5BL6 WT mice, CXCR5 -/- KO mice, and CXCR5 -/- .NRF2 -/- DKO mice retinal flatmounts were used for staining and quantification. (A-D) Retinal flatmounts stained with peanut agglutinin (PNA) lectin (A), MAP2 (B), cleaved caspase 3 at the ganglion cell layer (C) and the photoreceptor cell layer depth (D). The quantitative results of PNA lectin (+) photoreceptors (E), MAP2 (+) retinal ganglion cells (F). Caspase 3 (+) ganglion cells layer (G), and Caspase 3 (+) photoreceptor layer depths (H). The values of cell numbers per 100 µm 2 were averaged from 4 retinal samples (n = 4). P values were denoted: n.s. P > 0.05; * P

Techniques Used: Mouse Assay, Staining

19) Product Images from "Antioxidants Improve the Viability of Stored Adult Retinal Pigment Epithelial-19 Cultures"

Article Title: Antioxidants Improve the Viability of Stored Adult Retinal Pigment Epithelial-19 Cultures

Journal: Ophthalmology and Therapy

doi: 10.1007/s40123-014-0024-9

ImageJ quantification of caspase-3 expression after storage. Bar chart demonstrating caspase-3 expression in cultured ARPE-19 cells stored at 16 °C for 7 days. The cells were stored in unsupplemented MEM (control) or MEM supplemented with DADLE (1 mM), capsazepine (1 μM), DHA (50 nM) or resveratrol (30 μM). Compared to the unsupplemented MEM group, caspase-3 expression was significantly decreased for both DHA (50 nM) and resveratrol (30 μM) when assessed by ImageJ. * P
Figure Legend Snippet: ImageJ quantification of caspase-3 expression after storage. Bar chart demonstrating caspase-3 expression in cultured ARPE-19 cells stored at 16 °C for 7 days. The cells were stored in unsupplemented MEM (control) or MEM supplemented with DADLE (1 mM), capsazepine (1 μM), DHA (50 nM) or resveratrol (30 μM). Compared to the unsupplemented MEM group, caspase-3 expression was significantly decreased for both DHA (50 nM) and resveratrol (30 μM) when assessed by ImageJ. * P

Techniques Used: Expressing, Cell Culture

Photomicrographs showing the expression of RPE65 ( a , e ) and caspase-3 ( b , f ) in cultured ARPE-19 cells. Selections ( red ) around all cell nuclei ( white ) ( c , d ) were automatically created by ImageJ software. RPE65 ( e ) and caspase-3 ( f ) fluorescence intensity was then measured solely within the red selections, thereby normalizing for the number of cells in each image. Magnification ×200. ARPE-19 adult retinal pigment epithelium 19, Cy3 cyanine dye 3, DAPI 4′, 6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, RPE65 retinal pigment epithelium-specific protein 65 kDa
Figure Legend Snippet: Photomicrographs showing the expression of RPE65 ( a , e ) and caspase-3 ( b , f ) in cultured ARPE-19 cells. Selections ( red ) around all cell nuclei ( white ) ( c , d ) were automatically created by ImageJ software. RPE65 ( e ) and caspase-3 ( f ) fluorescence intensity was then measured solely within the red selections, thereby normalizing for the number of cells in each image. Magnification ×200. ARPE-19 adult retinal pigment epithelium 19, Cy3 cyanine dye 3, DAPI 4′, 6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, RPE65 retinal pigment epithelium-specific protein 65 kDa

Techniques Used: Expressing, Cell Culture, Software, Fluorescence

20) Product Images from "Transcriptional coactivator with PDZ‐binding motif is required to sustain testicular function on aging"

Article Title: Transcriptional coactivator with PDZ‐binding motif is required to sustain testicular function on aging

Journal: Aging Cell

doi: 10.1111/acel.12631

Increased level of senescence and damage markers in TAZ KO testicles. Testes were collected from WT and KO mice (12 months old, n = 6/group). (a) Immunoblotting analysis with antibodies against p21, p19, p16, γH2 AX , catalase, CDK 4, PCNA , and actin. (b) Testis tissue sections were analyzed by immunohistochemistry using antibodies against p21, p19, γH2 AX , and catalase. Representative images are shown. Arrows and arrowheads indicate positive cells in spermatogonial and the interstitial Leydig cells, respectively. (c) Quantitative image analysis of p21, p19, γH2 AX , and catalase from 10 images of six mice using ImageJ software (c). ** P
Figure Legend Snippet: Increased level of senescence and damage markers in TAZ KO testicles. Testes were collected from WT and KO mice (12 months old, n = 6/group). (a) Immunoblotting analysis with antibodies against p21, p19, p16, γH2 AX , catalase, CDK 4, PCNA , and actin. (b) Testis tissue sections were analyzed by immunohistochemistry using antibodies against p21, p19, γH2 AX , and catalase. Representative images are shown. Arrows and arrowheads indicate positive cells in spermatogonial and the interstitial Leydig cells, respectively. (c) Quantitative image analysis of p21, p19, γH2 AX , and catalase from 10 images of six mice using ImageJ software (c). ** P

Techniques Used: Mouse Assay, Immunohistochemistry, Software

Suppression of p21 expression and senescence by TAZ . (a, b) Primary testicular cells isolated WT and TAZ KO mice (12 months old, n = 6 each group) were maintained for three passages. Cells were subjected to a SA ‐βgal staining (a) and β‐galactosidase activity assay (b). Data are mean ± SD of six experiments. *** P
Figure Legend Snippet: Suppression of p21 expression and senescence by TAZ . (a, b) Primary testicular cells isolated WT and TAZ KO mice (12 months old, n = 6 each group) were maintained for three passages. Cells were subjected to a SA ‐βgal staining (a) and β‐galactosidase activity assay (b). Data are mean ± SD of six experiments. *** P

Techniques Used: Expressing, Isolation, Mouse Assay, Staining, Activity Assay

Suppression of p53 activity by TAZ via direct interaction. (a) Immunoblotting analysis of p53 in the testis of WT and KO mice (12 months old, n = 6/group). (b–d) 293T cells were transfected with the reporter gene either PG 13‐luc (b) or p21‐luc (d) and/or TAZ expression vector. Expression of p53 and TAZ was detected by immunoblotting analysis (c). The reporter activity was determined after normalization with β‐galactosidase activity and given as the fold induction compared to mock control. (e, f) 293T cells were transfected with Flag‐tagged p53 and Myc‐tagged TAZ expression vector. Whole cell extracts ( WCE ) were harvested and used for immunoprecipitation with anti‐Flag antibody, followed by Immunoblot analysis (e). Or cell extracts were incubated with biotinylated p21 promoter, followed by precipitation and immunoblotting (f). The data in panel b and d indicate the mean ± SD of three independent experiments. *** P
Figure Legend Snippet: Suppression of p53 activity by TAZ via direct interaction. (a) Immunoblotting analysis of p53 in the testis of WT and KO mice (12 months old, n = 6/group). (b–d) 293T cells were transfected with the reporter gene either PG 13‐luc (b) or p21‐luc (d) and/or TAZ expression vector. Expression of p53 and TAZ was detected by immunoblotting analysis (c). The reporter activity was determined after normalization with β‐galactosidase activity and given as the fold induction compared to mock control. (e, f) 293T cells were transfected with Flag‐tagged p53 and Myc‐tagged TAZ expression vector. Whole cell extracts ( WCE ) were harvested and used for immunoprecipitation with anti‐Flag antibody, followed by Immunoblot analysis (e). Or cell extracts were incubated with biotinylated p21 promoter, followed by precipitation and immunoblotting (f). The data in panel b and d indicate the mean ± SD of three independent experiments. *** P

Techniques Used: Activity Assay, Mouse Assay, Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Incubation

21) Product Images from "Postsynaptic density protein 95 (PSD-95) is transported by KIF5 to dendritic regions"

Article Title: Postsynaptic density protein 95 (PSD-95) is transported by KIF5 to dendritic regions

Journal: Molecular Brain

doi: 10.1186/s13041-019-0520-x

ADPDZ3 expression reduces the level of PSD-95 in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using monoclonal anti-PSD-antibody and Cy3-conjugated anti-mouse IgG antibody or polyclonal anti-synapsin I antibody and Alexa Fluor® 647 anti-rabbit IgG antibody. They were then visualized using confocal microscopy. a Representative images of expressed neurons. Arrows indicate analyzed dendrites. Scale bar: 20 μm. b , c ADPDZ3 expression reduced the number of PSD-95 particles in the dendrites (GFP: 100.00% ± 6.08%, n = 32, 1766 μm; ADPDZ3: 82.27% ± 4.03%, n = 35, 1973 μm; * P
Figure Legend Snippet: ADPDZ3 expression reduces the level of PSD-95 in dendrites. Cultured rat hippocampal neurons were infected with Sindbis viruses encoding GFP or GFP-ADPDZ3 and incubated for 9 h to allow expression. The cultures were then subjected to immunostaining using monoclonal anti-PSD-antibody and Cy3-conjugated anti-mouse IgG antibody or polyclonal anti-synapsin I antibody and Alexa Fluor® 647 anti-rabbit IgG antibody. They were then visualized using confocal microscopy. a Representative images of expressed neurons. Arrows indicate analyzed dendrites. Scale bar: 20 μm. b , c ADPDZ3 expression reduced the number of PSD-95 particles in the dendrites (GFP: 100.00% ± 6.08%, n = 32, 1766 μm; ADPDZ3: 82.27% ± 4.03%, n = 35, 1973 μm; * P

Techniques Used: Expressing, Cell Culture, Infection, Incubation, Immunostaining, Confocal Microscopy

Complexes of PSD-95-KIF5 colocalized with GluA1 particles in dendrites. Cultured hippocampal neurons were transfected with PSD-95-GFP constructs and incubated for days. The cultures were immunostained with monoclonal anti-PSD-95 antibody and polyclonal GluA1 antibody; they were subsequently stained with C3-conjugated anti-mouse IgG and Alexa−Fluor® 647 anti-rabbit IgG antibody. a Representative images of immunostaining in the first row. Each image was merged to show colocalization in the second row. A colocalized image of PSD-95 and KIF5 was collated with the image of GluA1. The colocalized points appeared white. Scale bar: 20 μm. b Boxed dendrites were enlarged to see the colocalization of GluA1 with the complex of PSD-95-KIF5A. c Staufen expression increased the association of PSD-95 and KIF5A. HA-PSD-95 and FLAG- KIF5A were cotransfected with 1, 2, or 3 μg of Myc-Staufen, or without Myc-Staufen as a control. After immunoprecipitation using anti-FLAG antibody or mouse IgG, immunoprecipitates were analyzed by Western blotting using anti-HA antibody. The lower panel shows expression of each group. d Quantified data of Western blot analyses (0 μg of Staufen: 100.0% ± 0.00%, n = 3; 1 μg of Staufen: 141.6% ± 16.96%, n = 3; 2 μg of Staufen: 192.3% ± 6.59%; 3 μg of Staufen; 274.4% ± 42.30%, n = 3). N values indicate the number of independent experiments. e Schematic diagram showing GluA1-containing vesicle transport mediated by PSD-95-KIF5A complex in dendrites. TARP: transmembrane AMPA receptor regulatory protein
Figure Legend Snippet: Complexes of PSD-95-KIF5 colocalized with GluA1 particles in dendrites. Cultured hippocampal neurons were transfected with PSD-95-GFP constructs and incubated for days. The cultures were immunostained with monoclonal anti-PSD-95 antibody and polyclonal GluA1 antibody; they were subsequently stained with C3-conjugated anti-mouse IgG and Alexa−Fluor® 647 anti-rabbit IgG antibody. a Representative images of immunostaining in the first row. Each image was merged to show colocalization in the second row. A colocalized image of PSD-95 and KIF5 was collated with the image of GluA1. The colocalized points appeared white. Scale bar: 20 μm. b Boxed dendrites were enlarged to see the colocalization of GluA1 with the complex of PSD-95-KIF5A. c Staufen expression increased the association of PSD-95 and KIF5A. HA-PSD-95 and FLAG- KIF5A were cotransfected with 1, 2, or 3 μg of Myc-Staufen, or without Myc-Staufen as a control. After immunoprecipitation using anti-FLAG antibody or mouse IgG, immunoprecipitates were analyzed by Western blotting using anti-HA antibody. The lower panel shows expression of each group. d Quantified data of Western blot analyses (0 μg of Staufen: 100.0% ± 0.00%, n = 3; 1 μg of Staufen: 141.6% ± 16.96%, n = 3; 2 μg of Staufen: 192.3% ± 6.59%; 3 μg of Staufen; 274.4% ± 42.30%, n = 3). N values indicate the number of independent experiments. e Schematic diagram showing GluA1-containing vesicle transport mediated by PSD-95-KIF5A complex in dendrites. TARP: transmembrane AMPA receptor regulatory protein

Techniques Used: Cell Culture, Transfection, Construct, Incubation, Staining, Immunostaining, Expressing, Immunoprecipitation, Western Blot

PSD-95 is colocalized with KIF5. a Representative images of immunostaining. Scale bar: 20 μm. Cultured neurons were immunostained with polyclonal anti-PSD-95 antibody and monoclonal anti-KIF5 antibody (H2), followed by Alexa Fluor® 488 anti-rabbit IgG antibody for PSD-95 and Cy3-conjugated anti-mouse IgG antibody for KIF5. b Results of co-localization analysis. 53.17% ± 3.86% ( n = 19 dendrites, 1354 μm) of KIF5-immunopositve puncta are colocalized with PSD-95-immunopositve puncta and 62.55% ± 1.69% (n = 19 dendrites, 1354 μm) of PSD-95-immunopositive puncta are colocalized with KIF5-immunopositve puncta. c Results of proximity ligation assay. Cultured neurons were infected with Sindbis viruses encoding GFP and subjected to PLA. Red dots in PLA indicate an interaction between the two proteins. Scale bar: 20 μm. d Results of IP assays using rat brain lysates. In total 500 μg of rat brain lysate was used in the IP assays using 3 μg monoclonal anti-KIF5 antibodies or polyclonal PSD-95 antibodies. These were analyzed by Western blotting using the antibody indicated. Asterisks indicate interaction bands in the Western blot assays
Figure Legend Snippet: PSD-95 is colocalized with KIF5. a Representative images of immunostaining. Scale bar: 20 μm. Cultured neurons were immunostained with polyclonal anti-PSD-95 antibody and monoclonal anti-KIF5 antibody (H2), followed by Alexa Fluor® 488 anti-rabbit IgG antibody for PSD-95 and Cy3-conjugated anti-mouse IgG antibody for KIF5. b Results of co-localization analysis. 53.17% ± 3.86% ( n = 19 dendrites, 1354 μm) of KIF5-immunopositve puncta are colocalized with PSD-95-immunopositve puncta and 62.55% ± 1.69% (n = 19 dendrites, 1354 μm) of PSD-95-immunopositive puncta are colocalized with KIF5-immunopositve puncta. c Results of proximity ligation assay. Cultured neurons were infected with Sindbis viruses encoding GFP and subjected to PLA. Red dots in PLA indicate an interaction between the two proteins. Scale bar: 20 μm. d Results of IP assays using rat brain lysates. In total 500 μg of rat brain lysate was used in the IP assays using 3 μg monoclonal anti-KIF5 antibodies or polyclonal PSD-95 antibodies. These were analyzed by Western blotting using the antibody indicated. Asterisks indicate interaction bands in the Western blot assays

Techniques Used: Immunostaining, Cell Culture, Proximity Ligation Assay, Infection, Western Blot

22) Product Images from "The Absence of Indoleamine 2,3-Dioxygenase Inhibits Retinal Capillary Degeneration in Diabetic Mice"

Article Title: The Absence of Indoleamine 2,3-Dioxygenase Inhibits Retinal Capillary Degeneration in Diabetic Mice

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.17-22702

Characterization of isolated HREC. Isolated HREC were characterized by (A) PECAM-1 immunostaining and (B) an assay of DiL-Ac-LDL uptake. The images are representative of three independent experiments. Scale bar: 100 μm.
Figure Legend Snippet: Characterization of isolated HREC. Isolated HREC were characterized by (A) PECAM-1 immunostaining and (B) an assay of DiL-Ac-LDL uptake. The images are representative of three independent experiments. Scale bar: 100 μm.

Techniques Used: Isolation, Immunostaining

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    Cell Signaling Technology Inc 4 6 diamidino 2 phenylindole dapi solution solution cell signaling technology danvers ma usa
    Characterization of human primary prostate epithelial cell cultures. Notes: ( A ) Primary outgrowth of epithelial cells derived from a biopsy specimen. ( B ) Primary epithelial cell colony. ( C ) Characterization of epithelial cells: green fluorescence shows high-molecular-weight cytokeratins; blue fluorescence shows nuclei reactive with <t>4′,6-diamidino-2-phenylindole</t> <t>(DAPI).</t> No vimentin staining was observed in primary epithelial cell cultures (data not shown).
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    Characterization of human primary prostate epithelial cell cultures. Notes: ( A ) Primary outgrowth of epithelial cells derived from a biopsy specimen. ( B ) Primary epithelial cell colony. ( C ) Characterization of epithelial cells: green fluorescence shows high-molecular-weight cytokeratins; blue fluorescence shows nuclei reactive with 4′,6-diamidino-2-phenylindole (DAPI). No vimentin staining was observed in primary epithelial cell cultures (data not shown).

    Journal: Research and Reports in Urology

    Article Title: IL-8 secretion in primary cultures of prostate cells is associated with prostate cancer aggressiveness

    doi: 10.2147/RRU.S58643

    Figure Lengend Snippet: Characterization of human primary prostate epithelial cell cultures. Notes: ( A ) Primary outgrowth of epithelial cells derived from a biopsy specimen. ( B ) Primary epithelial cell colony. ( C ) Characterization of epithelial cells: green fluorescence shows high-molecular-weight cytokeratins; blue fluorescence shows nuclei reactive with 4′,6-diamidino-2-phenylindole (DAPI). No vimentin staining was observed in primary epithelial cell cultures (data not shown).

    Article Snippet: Slides were washed in PBS/0.025% Tween 20 three times for 5 minutes, after which the nuclei were stained by incubating the slides for 10 minutes in a 1 μg/mL 4′,6-diamidino-2-phenylindole (DAPI) solution (Cell Signaling Technology, Danvers, MA, USA [1 μg/mL in PBS]).

    Techniques: Derivative Assay, Fluorescence, Molecular Weight, Staining