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Santa Cruz Biotechnology 27hc 10um

a Schematic of cyp27a1 conversion of cholesterol to <t>27HC</t> in a macrophage. 27HC content ( b , n = 8, 6) and cyp27a1 expression ( c , n = 5) were evaluated in aortas from apoE +/+ and apoE −/− mice following receipt of standard chow or atherogenic diet, respectively. d C yp27a1 expression in circulating monocytes (n = 8) from apoE +/+ and apoE -/- mice administered diets as in ( b ). e , f Cyp27a1 expression was assessed in human atherosclerotic versus control arteries in two independent cohorts. Cohorts I ( e ) and II ( f ) contained n = 4 and 198, and 32 and 32 samples respectively. FPKM is fragments per kilobase of transcript per million mapped reads. Box-and-whisker plots are provided; central line denotes the median value, edges represent the upper and lower quartiles, and whiskers indicate minimum and maximum values. In ( b ), ( c ) and ( d ) values are expressed relative to findings in apoE +/+ . g–r Atherosclerosis and related parameters in apoE -/- background cyp27a1 fl/fl and cyp27a1 ΔMAC mice. Representative lipid-stained en face images of aortas ( g ) and lesion areas ( h ; percent of total surface area, n = 10 and 9). Representative lipid and haematoxylin-stained aortic root sections ( i ), and lesion areas ( j , n = 11 and 9). Representative immunohistochemistry images of Mac2 staining in aortic root ( k ; nuclei DAPI stained) and quantification of Mac2 positive staining expressed per unit lesion area ( l ; n = 8). m Monocyte recruitment to atherosclerotic lesions was evaluated by YG bead incorporation into circulating monocytes and quantification of beads per aortic root section 24 h later. N = 8. Immunohistochemical determinations of endothelial ICAM-1 ( n , p ) and VCAM-1 abundance ( o , q ) with representative images ( n , o ) and quantification ( p , q , n = 9 and 8). Arrows indicate endothelium. r Inflammatory gene expression in the aorta (n = 4). s Representative still images of leukocyte-endothelial cell adhesion in apoE -/- background cyp27a1 fl/fl and cyp27a1 ∆MAC mice. t Summary data for leukocyte velocity (n = 4). Scale bar equals 100 um ( i ) or 50 um ( k , n , o ). Except in ( e ) and ( f) , data are mean ± SEM. P values by two-sided Student’s t-test ( b – f , h , j , l , m , p , q , t ) or two-sided Mann-Whitney U-test ( r ) are shown. Source data are provided as a Source Data file.

27hc 10um, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 982 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 982 article reviews

27hc 10um - by Bioz Stars, 2026-10

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1) Product Images from "Macrophage-to-endothelial cell crosstalk by the cholesterol metabolite 27HC promotes atherosclerosis in male mice"

Article Title: Macrophage-to-endothelial cell crosstalk by the cholesterol metabolite 27HC promotes atherosclerosis in male mice

Journal: Nature Communications

doi: 10.1038/s41467-023-39586-z

a Schematic of cyp27a1 conversion of cholesterol to 27HC in a macrophage. 27HC content ( b , n = 8, 6) and cyp27a1 expression ( c , n = 5) were evaluated in aortas from apoE +/+ and apoE −/− mice following receipt of standard chow or atherogenic diet, respectively. d C yp27a1 expression in circulating monocytes (n = 8) from apoE +/+ and apoE -/- mice administered diets as in ( b ). e , f Cyp27a1 expression was assessed in human atherosclerotic versus control arteries in two independent cohorts. Cohorts I ( e ) and II ( f ) contained n = 4 and 198, and 32 and 32 samples respectively. FPKM is fragments per kilobase of transcript per million mapped reads. Box-and-whisker plots are provided; central line denotes the median value, edges represent the upper and lower quartiles, and whiskers indicate minimum and maximum values. In ( b ), ( c ) and ( d ) values are expressed relative to findings in apoE +/+ . g–r Atherosclerosis and related parameters in apoE -/- background cyp27a1 fl/fl and cyp27a1 ΔMAC mice. Representative lipid-stained en face images of aortas ( g ) and lesion areas ( h ; percent of total surface area, n = 10 and 9). Representative lipid and haematoxylin-stained aortic root sections ( i ), and lesion areas ( j , n = 11 and 9). Representative immunohistochemistry images of Mac2 staining in aortic root ( k ; nuclei DAPI stained) and quantification of Mac2 positive staining expressed per unit lesion area ( l ; n = 8). m Monocyte recruitment to atherosclerotic lesions was evaluated by YG bead incorporation into circulating monocytes and quantification of beads per aortic root section 24 h later. N = 8. Immunohistochemical determinations of endothelial ICAM-1 ( n , p ) and VCAM-1 abundance ( o , q ) with representative images ( n , o ) and quantification ( p , q , n = 9 and 8). Arrows indicate endothelium. r Inflammatory gene expression in the aorta (n = 4). s Representative still images of leukocyte-endothelial cell adhesion in apoE -/- background cyp27a1 fl/fl and cyp27a1 ∆MAC mice. t Summary data for leukocyte velocity (n = 4). Scale bar equals 100 um ( i ) or 50 um ( k , n , o ). Except in ( e ) and ( f) , data are mean ± SEM. P values by two-sided Student’s t-test ( b – f , h , j , l , m , p , q , t ) or two-sided Mann-Whitney U-test ( r ) are shown. Source data are provided as a Source Data file.

Figure Legend Snippet: a Schematic of cyp27a1 conversion of cholesterol to 27HC in a macrophage. 27HC content ( b , n = 8, 6) and cyp27a1 expression ( c , n = 5) were evaluated in aortas from apoE +/+ and apoE −/− mice following receipt of standard chow or atherogenic diet, respectively. d C yp27a1 expression in circulating monocytes (n = 8) from apoE +/+ and apoE -/- mice administered diets as in ( b ). e , f Cyp27a1 expression was assessed in human atherosclerotic versus control arteries in two independent cohorts. Cohorts I ( e ) and II ( f ) contained n = 4 and 198, and 32 and 32 samples respectively. FPKM is fragments per kilobase of transcript per million mapped reads. Box-and-whisker plots are provided; central line denotes the median value, edges represent the upper and lower quartiles, and whiskers indicate minimum and maximum values. In ( b ), ( c ) and ( d ) values are expressed relative to findings in apoE +/+ . g–r Atherosclerosis and related parameters in apoE -/- background cyp27a1 fl/fl and cyp27a1 ΔMAC mice. Representative lipid-stained en face images of aortas ( g ) and lesion areas ( h ; percent of total surface area, n = 10 and 9). Representative lipid and haematoxylin-stained aortic root sections ( i ), and lesion areas ( j , n = 11 and 9). Representative immunohistochemistry images of Mac2 staining in aortic root ( k ; nuclei DAPI stained) and quantification of Mac2 positive staining expressed per unit lesion area ( l ; n = 8). m Monocyte recruitment to atherosclerotic lesions was evaluated by YG bead incorporation into circulating monocytes and quantification of beads per aortic root section 24 h later. N = 8. Immunohistochemical determinations of endothelial ICAM-1 ( n , p ) and VCAM-1 abundance ( o , q ) with representative images ( n , o ) and quantification ( p , q , n = 9 and 8). Arrows indicate endothelium. r Inflammatory gene expression in the aorta (n = 4). s Representative still images of leukocyte-endothelial cell adhesion in apoE -/- background cyp27a1 fl/fl and cyp27a1 ∆MAC mice. t Summary data for leukocyte velocity (n = 4). Scale bar equals 100 um ( i ) or 50 um ( k , n , o ). Except in ( e ) and ( f) , data are mean ± SEM. P values by two-sided Student’s t-test ( b – f , h , j , l , m , p , q , t ) or two-sided Mann-Whitney U-test ( r ) are shown. Source data are provided as a Source Data file.

Techniques Used: Expressing, Control, Whisker Assay, Staining, Immunohistochemistry, Immunohistochemical staining, Gene Expression, MANN-WHITNEY

a Representative still images of leukocyte-endothelial cell adhesion evaluated by intravital microscopy in ERα fl/fl and ERα ∆EC mice injected with vehicle versus 27HC (20 mg/kg BW) daily for 3 days. b Summary data for leukocyte velocity in ( a ), n = 11-12. c – f ApoE -/- background ERα fl/fl and ERα ΔEC mice were placed on an atherogenic diet for 8 weeks and administered vehicle versus 27HC, and samples were obtained. Representative lipid-stained en face images of aortas ( c ), and lesion areas ( d , percent of total surface area). N = 13 and 10 for ERα fl/fl given vehicle versus 27HC, respectively, and n = 10 and 12 for ERα ΔEC given vehicle versus 27HC, respectively. Representative lipid and haematoxylin-stained aortic root sections ( e ), and lesion areas ( f , n = 10). g – o , Atherosclerosis and related parameters in bone marrow transplant experiments performed employing apoE -/- background cyp27a1 fl/fl versus cyp27a1 ΔMAC donors and apoE -/- background ERα fl/fl versus ERα ΔEC recipients. g Schematic of donors and recipients. Representative lipid-stained en face images of aortas ( h ) and lesion areas ( i , n = 11-12). Representative lipid and haematoxylin-stained aortic root sections ( j ), and lesion areas ( k , n = 11-12). Representative immunohistochemistry images of Mac2 staining in aortic root ( l ; nuclei DAPI stained) and quantification of Mac2 positive staining ( m ; expressed per unit lesion area, n = 8−10). Immunohistochemical determinations of endothelial ICAM-1 and VCAM-1 abundance with representative images ( n ) and quantification ( o , n = 8-9). Arrows indicate endothelium. Scale bar equals 100 um ( e , j ) or 50 um ( l , n ). Data are mean ± SEM. In ( b ) and ( d ), P values shown are by Krusakal-Wallis with Dunn’s post hoc testing. In all other graphs ( f , i , k , m , o ), P values shown are by ANOVA with Tukey’s post-hoc testing. Source data are provided as a Source Data file.

Figure Legend Snippet: a Representative still images of leukocyte-endothelial cell adhesion evaluated by intravital microscopy in ERα fl/fl and ERα ∆EC mice injected with vehicle versus 27HC (20 mg/kg BW) daily for 3 days. b Summary data for leukocyte velocity in ( a ), n = 11-12. c – f ApoE -/- background ERα fl/fl and ERα ΔEC mice were placed on an atherogenic diet for 8 weeks and administered vehicle versus 27HC, and samples were obtained. Representative lipid-stained en face images of aortas ( c ), and lesion areas ( d , percent of total surface area). N = 13 and 10 for ERα fl/fl given vehicle versus 27HC, respectively, and n = 10 and 12 for ERα ΔEC given vehicle versus 27HC, respectively. Representative lipid and haematoxylin-stained aortic root sections ( e ), and lesion areas ( f , n = 10). g – o , Atherosclerosis and related parameters in bone marrow transplant experiments performed employing apoE -/- background cyp27a1 fl/fl versus cyp27a1 ΔMAC donors and apoE -/- background ERα fl/fl versus ERα ΔEC recipients. g Schematic of donors and recipients. Representative lipid-stained en face images of aortas ( h ) and lesion areas ( i , n = 11-12). Representative lipid and haematoxylin-stained aortic root sections ( j ), and lesion areas ( k , n = 11-12). Representative immunohistochemistry images of Mac2 staining in aortic root ( l ; nuclei DAPI stained) and quantification of Mac2 positive staining ( m ; expressed per unit lesion area, n = 8−10). Immunohistochemical determinations of endothelial ICAM-1 and VCAM-1 abundance with representative images ( n ) and quantification ( o , n = 8-9). Arrows indicate endothelium. Scale bar equals 100 um ( e , j ) or 50 um ( l , n ). Data are mean ± SEM. In ( b ) and ( d ), P values shown are by Krusakal-Wallis with Dunn’s post hoc testing. In all other graphs ( f , i , k , m , o ), P values shown are by ANOVA with Tukey’s post-hoc testing. Source data are provided as a Source Data file.

Techniques Used: Intravital Microscopy, Injection, Staining, Immunohistochemistry, Immunohistochemical staining

a – c 27HC activation of NF-κB activity in HAEC via ERα. a , NF-κB luciferase activity was measured in HAEC treated with TNFα (10 ng/ml), E2 (10 −8 M),TNFα plus E2, or 27HC (20uM) for 6 h (n = 8). b , c Following transduction with control shRNA versus shRNA targeting ERα ( b , findings for 3 samples per group are shown), the experiments described in ( a ) were performed ( c , n = 8). d Venn diagrams of proteins identified by LC-MS/MS to have increased or decreased association with ERα upon E2 versus 27HC treatment in HAEC. Diagrams display intersects for common directional changes with the two ligands (upper panel), and opposing directional changes with the two ligands (lower two panels). e Co-immunoprecipitation (IP) of endogenous ERα and septin 11 in HAEC treated with vehicle versus E2 (left panel), or with vehicle versus 27HC (right panel) for 30 min. f HAEC were treated with 27HC, and IP of endogenous ERα was performed and immunoblotting was done for ERα and septin 11 (upper panel), or IP of septin 11 was performed followed by immunoblotting. Septin 11 expression in human atherosclerotic versus control arteries in two independent cohorts. Cohorts I ( g ) and II ( h ), which were analyzed by RNAseq and microarray, respectively, contained n = 4 and 201, and 32 and 32 samples respectively. FPKM is fragments per kilobase of transcript per million mapped reads. Box-and-whisker plots are provided in which the central line denotes the median value, the edges represent the upper and lower quartiles, and whiskers indicate the minimum and maximum values. Effect of vehicle versus 27HC on NF-κB activity in HAEC expressing or lacking ERα ( i , n = 8) or septin 11 ( j , n = 4). In ( a ), ( c ), ( i ) and ( j ), NF-κB activity is expressed relative to values with vehicle treatment. Data are mean ± SEM. In ( a ) and ( c ), P values shown are by Krusakal-Wallis with Dunn’s post hoc testing, and in ( g ) and ( h ) two-sided Student’s t tests were used. In ( i ) and ( j ), P values by ANOVA with Tukey’s post-hoc testing are shown. The findings in ( e ) and ( f ) were confirmed in two independent experiments. Source data are provided as a Source Data file.

Figure Legend Snippet: a – c 27HC activation of NF-κB activity in HAEC via ERα. a , NF-κB luciferase activity was measured in HAEC treated with TNFα (10 ng/ml), E2 (10 −8 M),TNFα plus E2, or 27HC (20uM) for 6 h (n = 8). b , c Following transduction with control shRNA versus shRNA targeting ERα ( b , findings for 3 samples per group are shown), the experiments described in ( a ) were performed ( c , n = 8). d Venn diagrams of proteins identified by LC-MS/MS to have increased or decreased association with ERα upon E2 versus 27HC treatment in HAEC. Diagrams display intersects for common directional changes with the two ligands (upper panel), and opposing directional changes with the two ligands (lower two panels). e Co-immunoprecipitation (IP) of endogenous ERα and septin 11 in HAEC treated with vehicle versus E2 (left panel), or with vehicle versus 27HC (right panel) for 30 min. f HAEC were treated with 27HC, and IP of endogenous ERα was performed and immunoblotting was done for ERα and septin 11 (upper panel), or IP of septin 11 was performed followed by immunoblotting. Septin 11 expression in human atherosclerotic versus control arteries in two independent cohorts. Cohorts I ( g ) and II ( h ), which were analyzed by RNAseq and microarray, respectively, contained n = 4 and 201, and 32 and 32 samples respectively. FPKM is fragments per kilobase of transcript per million mapped reads. Box-and-whisker plots are provided in which the central line denotes the median value, the edges represent the upper and lower quartiles, and whiskers indicate the minimum and maximum values. Effect of vehicle versus 27HC on NF-κB activity in HAEC expressing or lacking ERα ( i , n = 8) or septin 11 ( j , n = 4). In ( a ), ( c ), ( i ) and ( j ), NF-κB activity is expressed relative to values with vehicle treatment. Data are mean ± SEM. In ( a ) and ( c ), P values shown are by Krusakal-Wallis with Dunn’s post hoc testing, and in ( g ) and ( h ) two-sided Student’s t tests were used. In ( i ) and ( j ), P values by ANOVA with Tukey’s post-hoc testing are shown. The findings in ( e ) and ( f ) were confirmed in two independent experiments. Source data are provided as a Source Data file.

Techniques Used: Activation Assay, Activity Assay, Luciferase, Transduction, Control, shRNA, Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Western Blot, Expressing, Microarray, Whisker Assay

a HAEC were treated with 27HC and immunoblotting was performed to detect Jnk Thr183/Tyr185 phosphorylation and total Jnk. b Following transfection of HAEC with control shRNA or shRNA targeting Jnk1 (upper panel, findings for 2 samples/group are shown), the effects of vehicle versus 27HC on NF-κB activity were determined (lower panel, n = 8). Following transduction of HAEC with control shRNA or shRNA targeting ERα ( c ) or septin 11 ( d ), HAEC were treated with 27HC, and immunoblotting was performed to detect activating phosphorylation of MLK3, MKK7 and Jnk, and total MLK3, MKK7 and Jnk. HAEC were treated with 27HC for 0 to 10 min, MKK7 was IP’d and immunoblotting was performed for MKK7 and GADD45β ( e ), or septin 11 was IP’d and immunoblotting was done for MKK7 and septin 11 ( f ). g In pull-down experiments a complex between HA-tagged bound MKK7 and GADD45β was formed. Wild-type septin 11 (WT) or septin 11 deletion mutants lacking AA184-192 (septin 11-∆G4) or AA38-304 (septin 11-∆GTP) were added, and protein associations were evaluated by immunoblotting. h , i Requirement for septin 11 interaction with MKK7 in 27HC action was evaluated in HAEC in which endogenous septin 11 was knocked down by shRNA, and reconstitution was performed with either wild-type septin 11, septin 11-∆G4, or septin 11-∆GTP. ( h ) Cells were treated with vehicle or 27HC, and immunoblotting was performed to detect activating phosphorylation of MKK7 and Jnk, and total MKK7 and Jnk. ( i ) In the same study groups as in ( h ), NF-κB activity was evaluated in cells treated with vehicle or 27HC. n = 8. In ( b ) and ( i ), NF-κB activity is expressed relative to values with vehicle treatment. Data are mean ± SEM. In ( b ) and ( i ), P values by ANOVA with Tukey’s post-hoc testing are shown. The findings in ( a ) and ( c )–( h ) were confirmed in two independent experiments. Source data are provided as a Source Data file.

Figure Legend Snippet: a HAEC were treated with 27HC and immunoblotting was performed to detect Jnk Thr183/Tyr185 phosphorylation and total Jnk. b Following transfection of HAEC with control shRNA or shRNA targeting Jnk1 (upper panel, findings for 2 samples/group are shown), the effects of vehicle versus 27HC on NF-κB activity were determined (lower panel, n = 8). Following transduction of HAEC with control shRNA or shRNA targeting ERα ( c ) or septin 11 ( d ), HAEC were treated with 27HC, and immunoblotting was performed to detect activating phosphorylation of MLK3, MKK7 and Jnk, and total MLK3, MKK7 and Jnk. HAEC were treated with 27HC for 0 to 10 min, MKK7 was IP’d and immunoblotting was performed for MKK7 and GADD45β ( e ), or septin 11 was IP’d and immunoblotting was done for MKK7 and septin 11 ( f ). g In pull-down experiments a complex between HA-tagged bound MKK7 and GADD45β was formed. Wild-type septin 11 (WT) or septin 11 deletion mutants lacking AA184-192 (septin 11-∆G4) or AA38-304 (septin 11-∆GTP) were added, and protein associations were evaluated by immunoblotting. h , i Requirement for septin 11 interaction with MKK7 in 27HC action was evaluated in HAEC in which endogenous septin 11 was knocked down by shRNA, and reconstitution was performed with either wild-type septin 11, septin 11-∆G4, or septin 11-∆GTP. ( h ) Cells were treated with vehicle or 27HC, and immunoblotting was performed to detect activating phosphorylation of MKK7 and Jnk, and total MKK7 and Jnk. ( i ) In the same study groups as in ( h ), NF-κB activity was evaluated in cells treated with vehicle or 27HC. n = 8. In ( b ) and ( i ), NF-κB activity is expressed relative to values with vehicle treatment. Data are mean ± SEM. In ( b ) and ( i ), P values by ANOVA with Tukey’s post-hoc testing are shown. The findings in ( a ) and ( c )–( h ) were confirmed in two independent experiments. Source data are provided as a Source Data file.

Techniques Used: Western Blot, Phospho-proteomics, Transfection, Control, shRNA, Activity Assay, Transduction

a–d ApoE -/- mice were placed on an atherogenic diet for 8 weeks, receiving vehicle or the cyp27a1 inhibitor GW273297X (GW) during the last 4 weeks, and atherosclerotic lesions were evaluated. Representative lipid-stained en face images of aortas ( a ) and lesion areas ( b ; percent of total surface area, n = 14). Representative lipid and haematoxylin-stained aortic root sections ( c ), and lesion areas ( d , n = 10). e – h In parallel experiments, apoE -/- mice received vehicle, cyp27a1 inhibitor GW, or GW plus exogenous 27HC during the last 4 weeks of an 8 week period on an atherogenic diet. Representative lipid-stained en face images of aortas ( e ) and lesion areas ( f , n = 9-13). Representative lipid and haematoxylin-stained aortic root sections ( g ), and lesion areas ( h , n = 8-9). i – l The effect of vehicle versus GW was determined in apoE -/- background cyp27a1 fl/fl and cyp27a1 ΔMAC mice. Representative lipid-stained en face images of aortas ( i ) and lesion areas ( j , n = 9-12). Representative lipid and haematoxylin-stained aortic root sections ( k ), and lesion areas ( l , n = 9-12). Effect of cyp27a1 inhibitor on abundance of endothelial ICAM-1 ( m, n ) and VCAM-1 abundance ( o , p ) with representative images ( m , o ) and quantification ( n , p , n = 8-9). q Effect of cyp27a1 inhibitor on monocyte recruitment, evaluated by YG bead incorporation into circulating monocytes and quantification of beads per aortic root section 24 h later. The quantitation was corrected for monocyte labeling efficiency. N = 7-8. r , s Effect of cyp27a1 inhibitor on lesion macrophage accumulation. Representative immunohistochemistry images of Mac2 staining in aortic root ( r ; nuclei DAPI stained) and quantification of Mac2 positive staining ( s ; expressed per unit lesion area, n = 8-9). Scale bar equals 100 um ( c , g , k ) or 50 um ( m , o , r ). Data are mean ± SEM, P values by two-sided Student’s t test ( b , d , n , p , q and s ), by Krusakal-Wallis with Dunn’s post hoc testing ( f ), or by ANOVA with Tukey post-hoc testing ( h , j and l ) are shown. Source data are provided as a Source Data file.

Figure Legend Snippet: a–d ApoE -/- mice were placed on an atherogenic diet for 8 weeks, receiving vehicle or the cyp27a1 inhibitor GW273297X (GW) during the last 4 weeks, and atherosclerotic lesions were evaluated. Representative lipid-stained en face images of aortas ( a ) and lesion areas ( b ; percent of total surface area, n = 14). Representative lipid and haematoxylin-stained aortic root sections ( c ), and lesion areas ( d , n = 10). e – h In parallel experiments, apoE -/- mice received vehicle, cyp27a1 inhibitor GW, or GW plus exogenous 27HC during the last 4 weeks of an 8 week period on an atherogenic diet. Representative lipid-stained en face images of aortas ( e ) and lesion areas ( f , n = 9-13). Representative lipid and haematoxylin-stained aortic root sections ( g ), and lesion areas ( h , n = 8-9). i – l The effect of vehicle versus GW was determined in apoE -/- background cyp27a1 fl/fl and cyp27a1 ΔMAC mice. Representative lipid-stained en face images of aortas ( i ) and lesion areas ( j , n = 9-12). Representative lipid and haematoxylin-stained aortic root sections ( k ), and lesion areas ( l , n = 9-12). Effect of cyp27a1 inhibitor on abundance of endothelial ICAM-1 ( m, n ) and VCAM-1 abundance ( o , p ) with representative images ( m , o ) and quantification ( n , p , n = 8-9). q Effect of cyp27a1 inhibitor on monocyte recruitment, evaluated by YG bead incorporation into circulating monocytes and quantification of beads per aortic root section 24 h later. The quantitation was corrected for monocyte labeling efficiency. N = 7-8. r , s Effect of cyp27a1 inhibitor on lesion macrophage accumulation. Representative immunohistochemistry images of Mac2 staining in aortic root ( r ; nuclei DAPI stained) and quantification of Mac2 positive staining ( s ; expressed per unit lesion area, n = 8-9). Scale bar equals 100 um ( c , g , k ) or 50 um ( m , o , r ). Data are mean ± SEM, P values by two-sided Student’s t test ( b , d , n , p , q and s ), by Krusakal-Wallis with Dunn’s post hoc testing ( f ), or by ANOVA with Tukey post-hoc testing ( h , j and l ) are shown. Source data are provided as a Source Data file.

Techniques Used: Staining, Quantitation Assay, Labeling, Immunohistochemistry

Virus:

Article Title: IFITM knockout DF1 cells produce higher influenza and newcastle disease viral yields: a proof of concept for avian origin cell-based vaccine production.
Article Snippet: Vaccines remain essential for the control of infectious diseases during poultry production, especially in high density systems.. Many of poultry vaccines are currently grown in embryonated chicken eggs (ECE) or egg derived primary cells.. These systems can be relatively costly and present a potential risk of supply during pandemics when demand for ECE can be high.

Infection:

Staining:

Article Title: The disruption of COPII vesicles activates HSF-1 through SEC-23.
Article Snippet: .. NIH3T3 cells were fixed in 4% (w/v) formaldehyde in phosphate-buffered saline for 15 min at room temperature and stained with anti-HSF1 (1:500, Cat# sc-13516, Santa Cruz) and anti-SEC23A (1:1000, Cat# PA5-28984, Invitrogen) antibodies overnight at 4oC. .. DNA was counterstained using DAPI (Cat# D8417, Sigma-Aldrich).

Article Title: E- and N-cadherin drive hepatic polarity and lumen elongation via opposing effects on RhoA activity
Article Snippet: .. For staining of Par3 and NuMA, cells were fixed in 100% methanol for 2 min at 4 °C followed by 1.8% formaldehyde (Santa Cruz Biotechnology, sc-203049A) for 10 min. ..

FACS:

Flow Cytometry:

Incubation:

Article Title: Hollow Spherical Nucleic Acids for Intracellular Gene Regulation Based Upon Biocompatible Silica Shells
Article Snippet: .. After overnight incubation, cells were fixed by 1% formaldehyde (Santa Cruz Biotechnology) in PBS followed by three washes with PBS. .. The fixed cells were then imaged with a Zeiss LSM 510 inverted laser scanning confocal microscope at 60x magnification.

Article Title: Anillin-dependent actin assembly at PML NBs protects genome stability
Article Snippet: .. 24 h after APH addition, cells grown on sterile glass cover slips were fixed in 3.7% formaldehyde/1× PBS for 10 min, permeabilised with 0.5% Triton X-100/1× PBS and incubated with one or two of the following primary antibodies: PML (Santa-Cruz sc-966), p-AKT-Ser473 (Cell Signalling 9271), p-AKT-Ser473 (Proteintech, 28731-1-A), 53BP1 (Thermo Scientific, MA5-26830), γ-H2AX (abcam, ab2893). ..

Sterility:

Transfection:

Article Title: Ameliorating Effect of Glehnia littoralis Extract on Periodontitis Through Regulation of 11β-Hydroxysteroid Dehydrogenase Type 1 in an Experimental Periodontitis Model.
Article Snippet: Cells were then rinsed twice with PBS and measured using the Incucyte Live-Cell Analysis System (Sartorius, Göttingen, Germany). .. After transfection, cells were fixed by treating with 4% formaldehyde solution for 30 min and then cultured overnight at 4 ◦C using 11β-HSD1 antibody (Santa Cruz Biotechnology Inc., cat. no. sc-518168, Dallas, TX, USA). .. Afterwards, the cells were fixed with 4% polyformaldehyde for 30 min and stained with 0.1% Alizarin Red S at room temperature for 1 h to stain the calcified nodules, and then observed under an optical microscope (Nikon, Tokyo, Japan) to photograph the calcified nodules.

Cell Culture:

other:

Article Title: Ameliorating Effect of Glehnia littoralis Extract on Periodontitis Through Regulation of 11β-Hydroxysteroid Dehydrogenase Type 1 in an Experimental Periodontitis Model
Article Snippet: To quantify the amount of calcium accumulation in the calcified nodules, the calcium was dissolved in 10% cetylpyridinium chloride (CPC) and the absorbance was measured at 560 nm using a multifunctional microplate reader (M1000 Pro, TECAN, Männedorf, Switzerland), referring to the method of a previously reported study [ ].

Review