Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: Ethanol’s Effects on Transient Receptor Potential Channel Expression in Brain Microvascular Endothelial Cells

doi: 10.1007/s11481-018-9796-3

Figure Lengend Snippet: Endothelial cell barrier permeability following treatment with a) different concentrations of EtOH or b) the TRPM7 antagonists, NS8593, CyPPA, or SKA-31. * p

Article Snippet: In the presence of 50 μM of each of the three blockers, NS8593, CyPPA, and SKA-31, the permeability of the BMVEC barrier significantly increased in 24 h ( and of Online Resource 1).

Techniques: Permeability

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: Long-term SKA-31 administration does not induce gross tissue damage. Panels A-C display H E histological staining of kidney sections (400x magnification) derived from vehicle treated young animals (A) and aged rats treated with either vehicle (B) or 10 mg/kg SKA-31 (C). BC: Bowman’s capsule, BS: Bowman’s Space, G: Glomerulus. H E staining of liver sections (400x magnification) from vehicle treated young animals, and vehicle and SKA-31 treated aged rats is presented in panels D, E and F, respectively. H: Hepatocyte, N: Nucleus, S: Sinusoid, PV: Portal Vein. Panels G-I illustrate H E staining of brain cerebellar sections (400x magnification) from vehicle treated young animals (G), and vehicle (H) and SKA-31 treated aged rats (I). GL: Granule cell Layer, PC: Purkinje Cells, ML: Molecular cell Layer. The scale bar displayed in the bottom right corner of each image represents 50 microns. Quantification of select histological parameters in kidney, liver and cerebellum is presented in panels J-Q, as follows: glomerular size relative to area of Bowman’s capsule (J), area of Bowman’s space relative to Bowman’s capsule (K), hepatocyte diameter (L), hepatocyte density per 5mm 2 (M), molecular cell layer thickness in the cerebellum (N), granule cell layer thickness in the cerebellum (O), cerebellar Purkinje cell density per 5mm 2 (P) and number of Purkinje cells per millimetre length of the molecular layer (Q). Structural measurements in stained sections were carried out using ImageJ software. White bars = vehicle-treated young rats, blue = vehicle treated aged rats, and red = SKA-31 treated aged rats. Statistical analyses were performed using one-way ANOVA and a Tukey’s post-hoc test; the asterisk indicates a statistically significant difference between the indicated groups, P

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques: Staining, Derivative Assay, Software

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: Long-term SKA-31 treatment does not promote a pro-inflammatory state .

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques:

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: SKA-31 treatment improves cardiovascular structure and function Panels A-D depict primary parameters (i.e. fractional shortening, ejection fraction, stroke volume and heart rate) of cardiac function, as determined by echocardiographic imaging (B-mode and M-mode) and analyses, in vehicle treated young animals (white), vehicle (blue) and SKA-31 treated (red) aged rats. Data are expressed as means ± S.D. The asterisk indicates a statistically difference between the indicated groups, as determined by ANOVA and a Tukey’s post-hoc test (n = 6 animals/group, P˂ 0.05). Panels E-G present H E staining (400x magnification) of the left ventricular free wall from young rats (E), vehicle (F) and SKA-31 treated aged rats (G). Scale bar in the bottom right corner of each image represents 50 microns; MC: myocyte. The H E stained sections in panels H-J (400x magnification) depict cross sections of thoracic aorta isolated from young animals (H), vehicle (I) and SKA-31 treated aged rats (J). Scale bar in the bottom right corner of each image represents 50 microns; TA: tunica adventitia, TM: tunica media, CT: connective tissue. Panels K and L quantify the size of individual LV myocytes (n ≥50 cells analyzed per tissue) and the density/number of LV myocytes per 5mm 2 of tissue area in the three treatment groups, respectively. Medial layer thickness of the thoracic aorta and the density/number of aortic myocytes per 5 mm 2 of tissue area are depicted in panels M and N, respectively. Colour coding as in panels A-D. Panels O-Q display representative picrosirius red-stained sections of the left ventricular (LV) free wall from young rats (O), vehicle (P) and SKA-31 treated aged rats (Q). Scale bar in the bottom right corner of each image represents 50 microns. The histogram in panel R quantifies the picrosirius red (PSR)-positive area in LV tissue from each group. Measurements of cell and tissue dimensions were performed using a manual tracing function in ImageJ. Statistical analysis was performed using one-way ANOVA, followed by a Tukey’s post-hoc test (n = 6 animals); * denotes a statistically significant difference between the indicated groups, P

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques: Imaging, Staining, Isolation

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: SKA-31 administration modulates the expression of key proteins in mesenteric arteries. . Panel I shows the mRNA levels of key signaling proteins in mesenteric arteries from SKA-31 treated aged rats (red bars), in comparison with vehicle treated aged rats (blue bars), as determined by qPCR analysis. Data in aged animals are normalized to the expression level of the same target mRNA detected in mesenteric arteries from vehicle treated young rats; data normalization was calculated using REST software. GAPDH was utilized as an internal reference mRNA in all determinations. The asterisk indicates a statistically significant difference (P

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Software

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: SKA-31 administration regulates the expression of calcineurin/NFAT signaling components in T cells Panel A displays representative FACS analysis of splenic lymphocytes (1×10 6 . Panel H displays the mRNA expression of key signaling proteins in splenic CD4+ T cells from vehicle (blue) and SKA-31 treated (red) aged rats. The qPCR results for each target have been normalized to the mRNA expression of the same gene observed in vehicle treated young animals; GAPDH was utilized as the internal reference gene for all qPCR analyses. Relative mRNA expression was calculated using REST software. An asterisk indicates a statistical difference compared with the vehicle treated aged rats, as determined by an unpaired Student’s t-test (P

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques: Expressing, FACS, Real-time Polymerase Chain Reaction, Software

Journal: Pharmacological research

Article Title: SKA-31, an activator of Ca2+-activated K+ channels, improves cardiovascular function in aging

doi: 10.1016/j.phrs.2019.104539

Figure Lengend Snippet: SKA-31 administration improves vascular reactivity in isolated arteries. Panels A-C display representative tracings showing vasodilatory responses evoked by acetylcholine (ACh, 0.3 μM), bradykinin (BK, 0.3 μM), sodium nitroprusside (SNP, 10 μM), adenosine (ADO, 10 μM) and pinacidil (Pina, 5 μM) in phenylephrine (PE) pre-constricted mesenteric arteries isolated from vehicle treated young (A), vehicle treated aged (B) and SKA-31 treated (10 mg/kg) aged male rats (C). The horizontal bars above and below the intraluminal diameter tracing indicate exposure to individual agents. In the second half of the protocol, arteries were acutely incubated with 0.3 μM SKA-31 to evaluate the potential augmentation of evoked vasodilation. (D) Histogram quantifying the average PE-induced contractile tone (% of maximal intraluminal diameter) in mesenteric arteries isolated from vehicle and SKA treated aged rats, and vehicle treated young rats. Each bar represents the calculated mean ± S.D. (n = 4–5 animals). (E) Histogram quantifying the inhibition of PE induced tone in isolated mesenteric arteries (n = 4–5 arteries/condition, mean ± S.D). The asterisk (*) denotes a statistically significant difference between the indicated groups, as determined by ANOVA, P

Article Snippet: SKA-31 was synthesized as previously described ( ) and dissolved in peanut oil (Sigma Aldrich) as a vehicle.

Techniques: Isolation, Incubation, Inhibition

Journal: PLoS ONE

Article Title: GAGE Cancer-Germline Antigens Are Recruited to the Nuclear Envelope by Germ Cell-Less (GCL)

doi: 10.1371/journal.pone.0045819

Figure Lengend Snippet: GAGE proteins are recruited to the nuclear envelope by GCL. A. Immunohistochemical double-staining of Myc-tagged human GCL and endogenous A-type lamins (lamins A/C) in transfected HeLa cells; GCL co-localized with lamins A/C at the nuclear envelope. B-E. HeLa cells were transiently transfected with GAGE12I (B), GCL-Myc (C), or GCL-Myc plus either GAGE12I (D) or GAGE1 (E). GCL localized at the nuclear envelope and recruited GAGE proteins from the nucleoplasm. F-G. Melanoma cell lines MZ2-MEL (F) and SK-MEL-31 (G), which express high levels of endogenous GAGE, were transfected to express GCL-Myc; immunostaining revealed GCL-mediated translocation of GAGE from the nucleoplasm to the nuclear envelope. H–K. Immunohistochemical analysis of endogenous GAGE proteins in human normal tissues and tumors revealed a dense GAGE signals near the nuclear envelope in cells of the fetal adrenal cortex (H), migrating primordial germ cells (I), breast carcinoma cells (J) and malignant melanoma cell (K) specimens. (GAGE = DAB/brown; Nuclei = Mayers hematoxylin/blue). Bars, 5 µm (A) and 10 µm (B-K).

Article Snippet: HeLa, HCT116, SK-MEL-31, HEK293 and A375 cells were purchased from ATCC (Manassas, VA, USA).

Techniques: Immunohistochemistry, Double Staining, Transfection, Immunostaining, Translocation Assay

Journal: PLoS ONE

Article Title: GAGE Cancer-Germline Antigens Are Recruited to the Nuclear Envelope by Germ Cell-Less (GCL)

doi: 10.1371/journal.pone.0045819

Figure Lengend Snippet: Estimated concentration of GAGE proteins in melanoma cells. A. Dot blots to estimate GAGE protein content in MZ2-MEL and SK-MEL-31 whole cell lysates by comparison to recombinant GAGE12I. Blots were probed with antibodies expected to recognize all GAGE family members; the number below each dot indicates the relative signal intensity. B. Relative quantification of GAGE proteins in nuclei and cytoplasm of MZ2-MEL and SK-MEL-31 cells based on immunostaining intensities of confocal images measured using ImageJ64 software.

Article Snippet: HeLa, HCT116, SK-MEL-31, HEK293 and A375 cells were purchased from ATCC (Manassas, VA, USA).

Techniques: Concentration Assay, Recombinant, Immunostaining, Software