Journal: PLoS Neglected Tropical Diseases

Article Title: The Cyclooctadepsipeptide Anthelmintic Emodepside Differentially Modulates Nematode, Insect and Human Calcium-Activated Potassium (SLO) Channel Alpha Subunits

doi: 10.1371/journal.pntd.0004062

Figure Lengend Snippet: A . Representative traces of currents recorded from HEK293 cells expressing eGFP alone (control), Ce SLO-1 or hum KCNMA1. Membrane potential was held at -60mV and stepped to between -100 and +90mV in 10mV increments for 50ms. Free [Ca 2+ ] in the internal solution was 300nM or 100μM as indicated. B . Current-voltage relationship for whole cell currents recorded from cells expressing eGFP alone (control), Ce SLO-1 or hum KCNMA1 at 300nM or 100μM free intracellular [Ca 2+ ]. Membrane potential was held at -60mV and stepped to between -100 and +90mV in 10mV increments for 50ms. Data points are the mean ± s.e.mean of ‘n’ cell recordings (shown in brackets). Currents recorded from Ce SLO-1 in the presence of 300nM Ca 2+ , Ce SLO-1 in the presence of 100μM Ca 2+ and hum KCNMA1 in the presence of 300nM Ca 2+ were significantly different from eGFP recorded in 300nM and 100μM Ca 2+ ; p<0.001, two-way ANOVA with Bonferroni post-hoc test.

Article Snippet: The alignment and identity of KCNMA1 isoform b with C . elegans slo-1a is described in Crisford et al, [ ]The transcript encoding this isoform (NM_002247) was available to purchase from OriGene Technologies, USA.

Techniques: Expressing, Control, Membrane

Journal: PLoS Neglected Tropical Diseases

Article Title: The Cyclooctadepsipeptide Anthelmintic Emodepside Differentially Modulates Nematode, Insect and Human Calcium-Activated Potassium (SLO) Channel Alpha Subunits

doi: 10.1371/journal.pntd.0004062

Figure Lengend Snippet: A . Representative whole cell currents recorded from HEK293 cells expressing either Ce SLO-1 or hum KCNMA1 before and after penitrem A (1μM) application. Membrane potential was held at -60mV and stepped to between -100 and +90mV in 10mV increments for 50ms. B . A time-course of steady state Ce SLO-1 and hum KCNMA1 currents at +70mV (stepped every 10s for 40ms from -60mV holding potential) before and during application of DMSO (0.01%) or penitrem A (1μM). The current response is shown as a percentage change from the mean of pre-drug current amplitude during the 0 to 2 min time period. An arrow indicates the time point at which the drug solution reached the chamber with the cells. Free intracellular [Ca 2+ ] was 100μM for Ce SLO-1 and 300nM for hum KCNMA1. Data points are the mean ± s.e.mean. p<0.0001 for penitrem A inhibited current compared to DMSO treatment, two-way ANOVA with Bonferroni post-hoc tests.

Article Snippet: The alignment and identity of KCNMA1 isoform b with C . elegans slo-1a is described in Crisford et al, [ ]The transcript encoding this isoform (NM_002247) was available to purchase from OriGene Technologies, USA.

Techniques: Expressing, Membrane

Journal: PLoS Neglected Tropical Diseases

Article Title: The Cyclooctadepsipeptide Anthelmintic Emodepside Differentially Modulates Nematode, Insect and Human Calcium-Activated Potassium (SLO) Channel Alpha Subunits

doi: 10.1371/journal.pntd.0004062

Figure Lengend Snippet: Time-course analysis of steady state hum KCNMA1 currents expressed in HEK293 cells at +70mV (stepped every 10s for 40ms from -60mV holding potential) before and during application of 0.01% DMSO or emodepside. The current response is shown as a percentage change from the mean of pre-drug current amplitude during the 0 to 2 min time period. Arrow indicates the time of emodepside application at A . 1nM B . 10nM and C . 100nM. Data points are the mean ± s.e.mean. 100nM emodepside had a biphasic effect on hum KCNMA1 currents eliciting a significant increase between 4 and 8 min application (p<0.05) and a significant decrease after 20 min application (p<0.001); two-way ANOVA with Bonferroni post-hoc tests.

Article Snippet: The alignment and identity of KCNMA1 isoform b with C . elegans slo-1a is described in Crisford et al, [ ]The transcript encoding this isoform (NM_002247) was available to purchase from OriGene Technologies, USA.

Techniques:

Journal: PLoS Neglected Tropical Diseases

Article Title: The Cyclooctadepsipeptide Anthelmintic Emodepside Differentially Modulates Nematode, Insect and Human Calcium-Activated Potassium (SLO) Channel Alpha Subunits

doi: 10.1371/journal.pntd.0004062

Figure Lengend Snippet: Analysis of the current-voltage relationship of the data sets shown in and Fig 7 for Ce SLO-1 and hum KCNMA1 whole cell currents expressed in HEK293 cells before and after treatment with emodepside (100nM). Ce SLO-1 currents were recorded before and 12 min after emodepside application. Hum KCNMA1 currents were recorded before and 5 min after emodepside application. Membrane potential was held at -60mV and stepped to between -100 and +90mV in 10mV increments for 50ms. Data points are the mean ± s.e.mean of n = 9 for Ce SLO-1 and n = 8 for hum KCNMA1. The amplitude of Ce SLO-1 currents in the presence of 100nM emodepside was significantly greater than currents recorded in the absence of emodepside between +20 and +90mV and for hum KCNMA1 was significantly greater than currents recorded in the absence of emodepside between +70 and +90mV; p<0.05; two-way ANOVA with Bonferroni post-hoc tests.

Article Snippet: The alignment and identity of KCNMA1 isoform b with C . elegans slo-1a is described in Crisford et al, [ ]The transcript encoding this isoform (NM_002247) was available to purchase from OriGene Technologies, USA.

Techniques: Membrane

Journal: BMC Neuroscience

Article Title: Ability of naringenin, a bioflavonoid, to activate M-type potassium current in motor neuron-like cells and to increase BK Ca -channel activity in HEK293T cells transfected with α- hSlo subunit

doi: 10.1186/s12868-014-0135-1

Figure Lengend Snippet: Stimulatory effect of NGEN on BK Ca -channel activity measured from α -hSlo -expressing HEK293T cells. (A) Original current traces showing the activity of BK Ca channels before (left) and after application (right) of 30 μM NGEN. Inside-out recordings were conducted with symmetrical K + concentration (145 mM). The potential was constantly held at +60 mV, and bath medium contained 0.1 μM Ca 2+ . Channel openings give an upward deflection in current. (B) Summary of the data showing effect of NGEN and NGEN plus verruculogen (Verr; 1 μM) on the probability of BK Ca -channel openings (mean ± SEM; n = 8-11 for each bar). * Significantly different from control. ** Significantly different from NGEN (30 μM) alone group.

Article Snippet: The pCMV6-XL4 vector containing human BK Ca -channel pore forming α-subunit cDNA (α- hSlo ; NM_002247.1) was obtained from Origene Technologies (Rockville, MD).

Techniques: Activity Assay, Expressing, Concentration Assay, Control

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: Primer sequences for RT-qPCR

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Sequencing

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a Images of skeletal muscle tissues acquired after immunohistochemistry assays. b Positive expression rate of KCNMA1. I/R ischemia-reperfusion, Sevo sevoflurane, KCNMA1 potassium calcium-activated channel subfamily M alpha 1; * p < 0.05 vs. the normal group; # p < 0.05 vs. the I/R group; scale bar = 25 µm; n = 10; measurement data are expressed as the mean ± standard deviation. Comparisons among multiple groups were performed by one-way ANOVA; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Immunohistochemistry, Expressing, Standard Deviation

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a Relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in the skeletal muscle tissues of mice detected by RT-qPCR. b Protein bands observed after Western blot analysis. c Relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle tissues of mice evaluated by Western blot analysis. I/R ischemia-reperfusion, Sevo sevoflurane, MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, VASP vasodilator-stimulated phosphoprotein, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, GAPDH glyceraldehyde-3-phosphate dehydrogenase; * p < 0.05 vs. the normal group; # p < 0.05 vs. the I/R group; n = 10; measurement data are expressed as the mean ± standard deviation. Comparisons among multiple groups were performed by one-way ANOVA; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a The predicted consequential pairing between MBNL1-AS1 and KCNMA1. b Relative luciferase activity in the NC, MBNL1-AS1 vector, and siRNA-MBNL1-AS1 groups determined by the dual-luciferase reporter gene assay. c RIP results in each group. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, si small interfering, NC negative control; * p < 0.05 vs. the NC group; measurement data are expressed as the mean ± standard deviation. Comparisons among multiple groups were performed by one-way ANOVA; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Luciferase, Activity Assay, Plasmid Preparation, Reporter Gene Assay, Negative Control, Standard Deviation

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a RT-qPCR measured the relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. b Protein bands measured by Western blot analysis. c Western blot analysis measured the relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, p-PKGII phosphorylated-PKGII, VASP vasodilator-stimulated phosphoprotein, p-VASP phosphorylated-VASP, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, si small interfering, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NC negative control; * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; measurement data are expressed as the mean ± standard deviation. Comparisons among multiple groups were done. Comparisons among multiple groups were performed by one-way ANOVA; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Western Blot, Negative Control, Control, Standard Deviation

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a Cell proliferation in each group as measured by MTT. b Cell proliferation in each group as measured by EdU. c Cell proliferation in each group as measured by EdU (×50). OD optical density, MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, si small interfering, NC negative control; * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; scale bar = 200 µm; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Negative Control, Control

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: a Cell cycle of skeletal muscle cells in seven groups as detected by flow cytometry. b Apoptosis of skeletal muscle cells in seven groups as detected by flow cytometry. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, si small interfering, NC negative control; * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; measurement data are expressed as the mean ± standard deviation. Comparisons among multiple groups were performed by one-way ANOVA; the experiment was repeated 3 times

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Flow Cytometry, Negative Control, Control, Standard Deviation

Journal: Experimental & Molecular Medicine

Article Title: RETRACTED ARTICLE: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: Apoptosis rate and cell cycle distribution of skeletal muscle cells among seven groups

Article Snippet: Next, the sections were incubated in 3% H 2 O 2 for 10 min and washed with distilled water, followed by antigen repair at high pressure for 90 s. After being cooled to room temperature, the sections were washed with phosphate buffer saline (PBS), blocked with 5% bovine serum albumin (BSA) for 30 min at 37 °C, and then incubated overnight at 4 °C with rabbit anti-mouse KCNMA1 primary antibodies (1:100, CSB-EP012077PI, Cusabio Biotech Co., Ltd., Wuhan, China).

Techniques: Control, Plasmid Preparation