Journal: BMC cancer

Article Title: Integrative machine learning frameworks to uncover specific protein signature in neuroendocrine cervical carcinoma.

doi: 10.1186/s12885-025-13454-z

Figure Lengend Snippet: Fig. 4 IHC staining of kNsDEPs in normal cervix specimens, paracancerous and tumor tissues of CSCC and NECC. (A-C) Representative IHC images of SCGN (A), CACYBP (B), and CAP2 (C) in the indicated specimens (n = 3). (D-F) Quantitative statistical analysis of protein levels of SCGN (D), CACYBP (E), and CAP2 (F) according to (A-C) and Figure S5. (n = 3) (*p < 0.05, **p < 0.01, ***p < 0.001; ns: not significant)

Article Snippet: The rabbit polyclonal antibody targeting CACYBP was obtained from Proteintech, USA (11745- 1-AP), with a primary antibody dilution of 1:200 for CACYBP.

Techniques: Immunohistochemistry

Journal: BMC cancer

Article Title: Integrative machine learning frameworks to uncover specific protein signature in neuroendocrine cervical carcinoma.

doi: 10.1186/s12885-025-13454-z

Figure Lengend Snippet: Fig. 5 Nomogram models based on kNsDEPs. (A) Nomogram model for NECC prediction based on SCGN, CAP2, and CACYBP. (B) The AUC score of (A). (C) Nomogram model for NECC prediction from TCGA based on SCGN, CAP2, and CACYBP. (D) The AUC score of (C)

Article Snippet: The rabbit polyclonal antibody targeting CACYBP was obtained from Proteintech, USA (11745- 1-AP), with a primary antibody dilution of 1:200 for CACYBP.

Techniques:

Journal: bioRxiv

Article Title: CFTR High Expresser BEST4+ cells are pH-sensing neuropod cells: new implications for intestinal physiology and Cystic Fibrosis disease

doi: 10.1101/2025.01.24.634747

Figure Lengend Snippet: All regions are mid villus or villus tip rat epithelium. (A) CHE cells expressing BEST4 (green) have long basal pseudopodia (arrow) that extend to submucosal neurons (arrowheads) expressing TUBB3 (red). (B) A CHE cell basal pseudopod expressing BEST4 (green) extends to a submucosal neuron expressing SYT3 (yellow). (C) BEST4+ (green) CHE cells and all villus epithelial cells express NFM (red) at the basolateral membrane. (D) A MEIS1+ (magenta) CHE cell highly expresses cytoplasmic and apical S100A6 (green). (E) A CHE cell is innervated by neurons expressing CHAT (red). A goblet cell (arrow) expressing CHAT (red) is found in close proximity to the CHE cell. (F) A MEIS1+ (magenta) CHE cell highly expresses cytoplasmic and apical GC-C (green). GC-C is also in scattered in vesicular structures in villus enterocytes (G) BEST4 (green) and TUBB3 (magenta) staining shows CHEs at the same plane and close proximity to enteric neurons. Scale bars: 10 µm.

Article Snippet: TUBB3 (#NB100-1612), TUBB2B (#NBP2-46250), SYT3 (#NBP1-19320), and S100A6 (#NB110-93274SS) were purchased from Novus Biologicals.

Techniques: Expressing, Membrane, Staining

Journal: Frontiers in Molecular Biosciences

Article Title: Aged Callus Skeletal Stem/Progenitor Cells Contain an Inflammatory Osteogenic Population With Increased IRF and NF-κB Pathways and Reduced Osteogenic Potential

doi: 10.3389/fmolb.2022.806528

Figure Lengend Snippet: Validation of increased CXCR2 high cells in the callus of aged mice with increased IRF and NF-κB pathways and reduced osteogenic potential. The tibial fracture was performed on 4-month- (young) and 21-month-old (aged) C57BL/6J male mice and fracture callus was harvested 10 days after the procedure. (A) Immunostaining with anti-CXCR2 antibody in callus sections for CXCR2+ cells. Images show numerous CXCR2+ cells (arrows) localize on the surface of woven bone. The percentage of CXCR2+ cells in the callus of young and aged mice was quantified by Image J software. n = 4 mice/young and 3 mice/aged. Data represent mean ± SD. Unpaired two-tailed t-test. * p < 0.05. WB: Woven bone. Scale bar = 1 mm. (B) Western blot to assess IRF-response protein IFITM1 and NF-κB-response protein S100A6 in young and aged callus tissues. (C) Violin plot of the expression level of Cxcr2 in cluster 1.1 and cluster 1.2. (D) CXCR2 low and CXCR2 high cells were isolated from callus-derived mesenchymal progenitor culture. (E) The expression levels of target genes for IRF and NF-κB were measured by RT-qPCR. n = 3 mice/group. Data represent mean ± SD. Unpaired two-tailed t-test. * p < 0.05. (F) CXCR2 low and CXCR2 high cells isolated from callus-derived mesenchymal progenitor culture were cultured in osteoblast inducing medium for 7 days and stained for ALP. The percentage area of ALP+ was measured in Image J software. Expression level of Alp and Runx2 was measured with RT-qPCR. Data represent mean ± SD. Unpaired two-tailed t -test. * p < 0.05.

Article Snippet: Proteins were loaded onto 15% SDS-PAGE gel and blotted with anti-IFITM1 Ab (Cell Signaling 13126, 1:500) and anti-S100A6 Ab (Cell Signaling 13162, 1:500).

Techniques: Biomarker Discovery, Immunostaining, Software, Two Tailed Test, Western Blot, Expressing, Isolation, Derivative Assay, Quantitative RT-PCR, Cell Culture, Staining

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: Expression of S100 family members is associated with EMT, and mesenchymal S100 proteins stimulate invasion of PDAC cells. a Immunoblot analysis of EMT-TFs, EMT markers and S100 proteins in a panel of PDAC cell lines. b Analysis of the transcription of ZEB1-regulated S100 genes in epithelial PDAC cells. BxPC-3 and SU.86.86 cell lines were transfected with the plasmid vectors expressing GFP-tagged ZEB1 or GFP control and cultured for 48 h. Bar charts show the expression of genes encoding S100 proteins and EMT markers analysed by qPCR. Data represent the mean of three replicate experiments ± StDev. c Invasion of mesenchymal and epithelial PDAC cell lines in zebrafish embryos. Cells were fluorescently DilC12-labelled (red), microinjected into the perivitelline cavities of zebrafish embryos, imaged and analysed 48 hpi as described in Methods. Merged phase contrast and fluorescence images are representative examples of zebrafish embryos with no invasion (upper image) or with PDAC cells that intravasate into the circulation (lower image). Minimum 10 fish per cell line were used in each experiment. Results are mean ± StDev of three independent experiments. d Mesenchymal S100 proteins, S100A4 and S100A6, contribute to the enhanced invasive potential of PDAC cells in vivo. siRNA-mediated knockdowns of mesenchymal S100 genes reduce the invasion of AsPC-1 and MIA PaCa-2 cells in zebrafish assay. Western blots show the extent of depletion of S100A4 and S100A6. Results are means ( n = 3 biological replicates; 10 fish in each experiment) ± StDev. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Control, Cell Culture, Fluorescence, In Vivo

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: IL-11/STAT3 and ZEB1 regulate expression of S100 proteins in a mutually independent mode. a Treatment with IL-11 does not induce an EMT in epithelial PDAC cells. BxPC-3 and SU.86.86 cells were treated with IL-11 or mock-treated and the expression of EMT-TFs and EMT markers was analysed by immunoblotting as indicated. b Ectopic expression of ZEB proteins does not affect STAT3 phosphorylation. Effect of ZEB proteins on S100A4, S100A6 and pSTAT3 expression levels was analysed in indicated epithelial cell lines. Note that ZEB proteins regulate S100A4 and S100A6 independently of STAT3. ( c ) Combined effects of ZEB1 and IL-11 on the expression of S100 proteins. ZEB1-expressing or non-expressing cells were treated with IL-11 or left untreated, and level of the indicated S100 proteins was analysed by immunoblotting. d Schematic representation of EMT-ZEB1 and IL-11/6-STAT3 pathways regulating expression of mesenchymal (S100A4 and S100A6) and an epithelial (S100A14) S100 proteins in PDAC cells. ZEB1 cooperates with the inflammatory signalling to induce S100A4 and S100A6. On the other hand, it abolishes an effect of IL-11/IL-6 on the expression of S100A14. Reduced expression of S100A14 further stimulates production of S100A4 and S100A6. This regulatory network drives the invasion of PDAC cells

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Expressing, Western Blot, Phospho-proteomics

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: S100 proteins are expressed in pancreatic lesions. a Examples of IHC analyses of the expression of S100 proteins in a series of PDAC samples ( n = 31). b Images exemplify co-expression of mesenchymal S100 proteins and E-caherin in parallel sections of a PDAC sample. c Bar charts illustrate the frequency of the expression of S100A4 and S100A6 proteins in different types of pancreatic lesions presented on TMA

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: Association between EMT markers and expression of S100 proteins in PDAC samples

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: IL-6/11-STAT3 signalling regulates expression of S100 proteins in PDAC cells. a IL-6 and IL-11 enhance expression levels of S100A4 and S100A6 proteins in PDAC cell lines. PDAC cells were exposed to the specified cytokines (200 ng/ml) for 48 h, and changes in protein expression levels were assessed by immunoblotting with indicated antibodies. b IL-11 enhances expression of S100 proteins belonging to both mesenchymal and epithelial groups. BxPC-3 and SU.86.86 cells were cultured in the presence of IL-11 for 48 h, and protein expression was analysed by immunoblotting. c STAT3 mediates the effect of IL-11 on the expression of S100 proteins in epithelial PDAC cell lines. Expression of S100 proteins was stimulated by IL-11 treatment for 48 h. A selective STAT3 inhibitor stattic was added 24 h before cells were harvested, and expression of S100 proteins was analysed by immunoblotting. d STAT3 controls expression of S100 proteins in mesenchymal cells, where their expression is intrinsically high. MIA PaCa-2 cells were treated with stattic for 24 h prior the immunoblot analysis with indicated antibodies was carried out. e Parallel sections from the same block containing a PDAC tissue were analysed by IHC. The images exemplify samples, in which S100A4, S100A6 and pSTAT3 proteins are co-expressed

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Expressing, Western Blot, Cell Culture, Blocking Assay

Journal: British Journal of Cancer

Article Title: ZEB1 and IL-6/11-STAT3 signalling cooperate to define invasive potential of pancreatic cancer cells via differential regulation of the expression of S100 proteins

doi: 10.1038/s41416-019-0483-9

Figure Lengend Snippet: Mesenchymal S100 proteins mediate IL-11/STAT3-driven invasion in zebrafish embryos. a IL-11 stimulates invasion of PDAC cells in STAT3-dependent manner. Epithelial PDAC cells were treated with the control siRNA, or siRNA targeting S100A4, S100A6, or STAT3, cultured in the presence or absence of IL-11 for 48 h, and analysed in zebrafish invasion assay. Expression of STAT3 and S100 proteins was analysed by immunoblotting. b BxPC-3 and SU.86.86 cells were treated with IL-11 alone or in combination with stattic, and their invasiveness was assessed in zebrafish embryos. c Mesenchymal MIA PaCa-2 cells were treated with stattic, or mock-treated, and invasion analysed in zebrafish embryos. a – c Results are mean ± StDev of biological replicates ( n = 3). Cell invasion was analysed in 10 fish in each experiment. * p < 0.05; ** p < 0.01; *** p < 0.001 (Student’s t -Test)

Article Snippet: We used the following primary antibodies raised against ZEB1 and S100A8, (both from Santa Cruz Biotechnology, Dallas, USA); S100A4 and S100A6 (both from Proteintech, Manchester, UK); S100A2, S100A9, S100A14, S100P, Tubulin and β-Actin (all from Sigma Aldrich); SNAIL1, SNAIL2, STAT3, pSTAT3, p65 (Cell Signaling Technology, Danvers, MA, USA); E-cadherin and P-cadherin (BD Bioscience, San Jose, CA, USA); S100A11 (R&D Systems, Minneapolis, MN, USA); TWIST1 (Abcam Cambridge, MA, USA); ZEB2 (in-house made ).

Techniques: Control, Cell Culture, Invasion Assay, Expressing, Western Blot

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 1. Localization and confirmation of in vitro transduction. Immunocytochemical staining of (A) exogenous GFP-tagged human S100A6 (green) in adenoviral-transduced neonatal cardiomyocytes showing perinuclear and cytoplasmic localization (magnification 960) and (B) endogenous rat S100A6 (green) in neonatal cardiomyocytes showing cytoplasmic and nuclear localization (magnification 960). C, Staining of endogenous S100A6 (top panel—green), S100A6-His fusion protein (middle panel—red) after cytoplasmic uptake of exogenously added S100A6 in the presence of 2 mmol/L Ca2+ after 1 hour, and colocalization (both cytoplasmic and nuclear) with endogenous S100A6 (bottom panel) (magnification 920). D, Expression of rat-S100A6, rat-S100B, rat-S100A1 and exogenous human/rat-S100A6 mRNA in S100A6-overexpressing and S100A6-knockdown neonatal cardiomyocytes by qRT-PCR. Data expressed as meanSEM, *P<0.05 vs nontransduced controls, N=7 for each. Overexpression and downregulation of S100A6 did not affect the expression of S100A1 or S100B in vitro. E, Western blot in S100A6- overexpressing and S100A6-knockdown cardiomyocytes compared to null-plasmid– and nontransduced controls. DAPI, 4ʹ,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GFP, green fluorescent protein; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: In Vitro, Transduction, Staining, Expressing, Knockdown, Quantitative RT-PCR, Over Expression, Western Blot, Plasmid Preparation, Real-time Polymerase Chain Reaction

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 2. Calcium transients and calcineurin (CaN) activity in vitro. A, Representative calcium transient imaging for nontransduced control, S100A6-knockdown (KD) and S100A6-overexpressing (OE) cardiomy- ocytes. B, The calcium release amplitude, the rate of calcium release and uptake in S100A6-overexpressing and S100A6-knockdown neonatal cardiomyocytes compared to nontransduced controls during sponta- neous action potentials (meanSEM, *P<0.05 vs nontransduced controls, N=7 for each). C, CaN phosphatase activity during normoxia and after H/R (24/6 hours) in S100A6-overexpressing and S100A6- knockdown neonatal cardiomyocytes compared to null-transduced and nontransduced controls and relative expression of CaN A and CaN B in S100A6-overexpressing, S100A6-knockdown, null-transduced, and nontransduced cardiomyocytes during normoxia and hypoxia/reoxygenation (meanSEM, *P<0.05 vs null- transduced and nontransduced controls, N=7 for each).

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: Activity Assay, In Vitro, Imaging, Control, Knockdown, Expressing

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 3. Effect of S100A6 on apoptosis after hypoxia/reoxygenation in vitro. S100A6 overexpression prevented hypoxia/reoxygenation (24/6 hours)-induced apoptosis in vitro, whereas S100A6-down- regulated cells showed greater apoptosis under normal conditions and after hypoxia/reoxygenation (24/ 6 hours). A, TUNEL staining in (i) nontransduced controls, (ii) S100A6-overexpressing, and (iii) S100A6- knockdown neonatal cardiomyocytes. Nuclei are stained by DAPI (blue) and apoptotic cells are rhodamine positive (red), 960. B, Rhodamine-positive apoptotic neonatal cardiomyocytes detected by TUNEL staining in S100A6-overexpressing and S100A6-knockdown neonatal cardiomyocytes compared to null-transduced and nontransduced cells (meanSEM, *P<0.05 vs null-transduced controls, N=10 for each). C, Caspase 3/ 7 activity in S100A6-overexpressing and S100A6-knockdown neonatal cardiomyocytes compared to null- transduced and nontransduced control cells after hypoxia/reoxygenation (24/6 hours) (meanSEM, *P<0.05 vs null-transduced controls, N=10 for each). DAPI, 4ʹ,6-diamidino-2-phenylindole; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: In Vitro, Over Expression, TUNEL Assay, Staining, Knockdown, Activity Assay, Control

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 4. Effect of UTMD of S100A6 on myocardial ischemia-reperfusion. Exogenous human S100A6 gene expression by q RT-PCR in various regions of myocardium (anterior LV, posterior LV, and RV) at (A) day 1, (B) day 3, and (C) day 28 post–ischemia/reperfusion (I/R) normalized to normal nonischemic control myocardium (meanSEM, *P<0.05 vs all other groups, N=7 per group). D, Mortality rate during/after myocardial I/ R in UTMD human-S100A6 plasmid-pretreated and UTMD empty-plasmid-pretreated rats compared to nontreated controls, N=29 per group. E, Representative M-mode echo image for (i) nontreated control, (ii) empty-plasmid-treated control, and (iii) UTMD S100A6-plasmid-treated. LV systolic function parameters ([F] ejection fraction and [G] fractional area change) following I/R in human-S100A6 plasmid-treated and empty plasmid-treated rats compared to nontreated controls (meanSEM, *P<0.05 vs nontreated and empty plasmid-treated controls, N=14-20 per group). H, Circumferential extent of LV akinesis following I/R in human-S100A6 plasmid-treated and empty plasmid-treated rats compared to nontreated control rats (meanSEM, *P<0.05 vs non-treated and empty plasmid-treated controls, N=14-20 per group). LV, left ventricle; RV, right ventricle; qRT-PCR, quantitative real-time polymerase chain reaction; UTMD, ultrasound-targeted microbubble destruction.

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Control, Plasmid Preparation, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 5. Cardiac hypertrophy data post–myocardial ischemia-reperfusion. A, Ratio of heart weight to tibia length at day 28 post-I/R (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=14-20 in each group). B, Ratio of heart weight to body weight at day 28 post-I/R (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=14-20 in each group). C, Representative H&E-stained images of noninfarct sections of LV myocardium at day 28 post-I/R in (i) control, (ii) empty plasmid, and (iii) S100A6 plasmid delivered rats. D, Cardiomyocyte size at day 28 post-I/R (magnification 9100) (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=50- 60 cells in each group). E, Relative expression of bMHC in the infarct and noninfarct regions of myocardium 28 days post-I/R (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=7 for all groups). F, Relative expression of ANF in the infarct and noninfarct regions of myocardium 28 days post-I/R (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=7 for all groups). ANF, atrial natriuretic factor; bMHC, b-myosin heavy chain; H&E, hematoxylin and eosin; I/R, ischemia-reperfusion.

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: Plasmid Preparation, Staining, Control, Expressing

Journal: Journal of the American Heart Association

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/jaha.116.004738

Figure Lengend Snippet: Figure 6. Infarct size and apoptosis data post–myocardial ischemia-reperfusion. A, Representative Evans blue–stained myocardial sections during ligation of the left anterior descending coronary artery, outlining the area at risk in nontreated controls, empty plasmid–treated controls, and human-S100A6 plasmid–treated rats. B, Representative TTC-stained myocardial cryosections 1 day post-I/R in nontreated controls, empty plasmid–treated controls, and human-S100A6 plasmid–treated rats. The nonviable/infarcted myocardium is white, and viable myocardium is red/pink. C, Representative MTC-stained myocardial cryosections 28 days post-I/R in nontreated controls, empty plasmid–treated controls, and human-S100A6 plasmid–treated rats. The collagen debris in the central compact necrotic region of the infarcted area is shown in blue, myocardial fibers in pink, and nuclei in purple. D, Risk area by Evans blue as percentage of the LV (meanSEM, N=6 per group). E, Nonviable myocardium/infarct area by TTC staining as percentage of the LV (meanSEM, *P<0.05 vs nontreated and empty plasmid–treated controls, N=9 per group). F, Infarct area/scar size by MTC staining as a percentage of the LV (meanSEM, *P<0.05 vs nontreated and empty plasmid– treated controls, N=8-9 per group). G, IHC staining of infarct region of myocardium 1 day post-I/R (sarcomeric a-actinin [red], nuclei [blue], and fluorescein isothiocyanate [FITC]-positive apoptotic nuclei [green]) (magnification 940). H, Data on apoptotic index in infarct and noninfarct regions in myocardial cryosections 1 day post-I/R in human-S100A6 plasmid–treated and empty plasmid–treated rats compared to nontreated controls (meanSEM, *P<0.05 vs nontreated and empty-treated controls, N=7-9 for each group). I, Caspase 3/7 activity in the infarct and noninfarct regions of myocardium 1 day post-I/R in S100A6-treated and empty plasmid–treated rats compared to nontreated controls (meanSEM, *P<0.05 vs nontreated and empty-treated controls, N=6 in each group). LV, left ventricle; MTC, Masson trichrome; TTC, 2,3,5-triphenyl-tetrazolium chloride.

Article Snippet: Membranes were incubated with rabbit monoclonal anti-S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1-95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti-rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B).

Techniques: Staining, Ligation, Plasmid Preparation, Immunohistochemistry, Activity Assay