Journal: Journal of Cardiovascular Medicine (Hagerstown, Md.)

Article Title: Empagliflozin ameliorates ventricular arrhythmias by inhibiting sympathetic remodeling via nerve growth factor/tyrosine kinase receptor A pathway inhibition

doi: 10.2459/JCM.0000000000001630

Figure Lengend Snippet: Empagliflozin treatment reduced sympathetic remodeling in myocardial infarction rats; the effect of the high-dose empagliflozin was more significant. (a) Presence of TH and GAP43 measured by IHC. (b) WB results of GAP43 and TH. The data are presented as the mean ± SD. n 1 (Sham group) = 10; n 2 (MI group) = 10; n 3 (low-EMPA group) = 11; n 4 (high-EMPA group) = 12. Two-way repeated measures ANOVA was performed. ∗ Means P < 0.05. ANOVA, analysis of variance; EMPA, empagliflozin; IHC, immunohistochemistry; SD, standard deviation; TH, tyrosine hydroxylase; WB, Western blotting.

Article Snippet: The sections were blocked with the 10% goat serum for 10 min at room temperature and then incubated with the anti-growth-associated protein 43 (GAP43) antibody (1 : 1000, ab75810, Abcam, Cambridge, UK) or the anti-tyrosine hydroxylase antibody (1 : 1000, ab112, Abcam) in 3% goat serum/1% BSA at 4 °C overnight.

Techniques: Immunohistochemistry, Standard Deviation, Western Blot

Journal: Life Science Alliance

Article Title: Nrg1 intracellular signaling regulates the development of interhemispheric callosal axons in mice

doi: 10.26508/lsa.202302250

Figure Lengend Snippet: (A) Western blot (WB) analysis of GAP43 protein levels in control (Nrg1 flox/flox ) and Nrg1-deficient (Nes-Cre; Nrg1 flox/flox ) neurons at DIV4. Tubulin levels are shown as a loading control. (A, B) Quantification of GAP43 protein levels from (A). GAP43 protein levels are normalized to tubulin. Data are represented as the mean ± SEM. Statistical significance was determined using an unpaired t test with Welch’s correction. Ctrl: n = 6; Nrg1 KO: n = 5, mice, from two litters (* P < 0.05). (C) Representative images and corresponding tracings of immunofluorescence staining in control, Nrg1-deficient (Nrg1 KO), and Nrg1 KO neurons expressing GAP43 (Nrg1 KO+GAP43) at DIV4. Scale bar, 50 μm. (D) Quantification of axonal length in control, Nrg1 KO, and Nrg1 KO+GAP43 neurons. Data are represented as the mean ± SEM. Statistical significance was determined using a one-way ANOVA test with Tukey’s post hoc test. Ctrl: n = 104; Nrg1 KO: n = 105; GAP43: n = 78, neurons from two independent experiments (** P < 0.01, *** P < 0.001). (E) Quantification of branch density in control, Nrg1 KO, and Nrg1 KO+GAP43 neurons. Data are represented as the mean ± SEM. Statistical significance was determined using a one-way ANOVA test with Tukey’s post hoc test. Ctrl: n = 93; Nrg1 KO: n = 98; GAP43: n = 73, neurons from two independent experiments. ns, not significant.

Article Snippet: Proteins from gels were transferred onto PVDF membranes (03010040001; Roche), and they were incubated with the primary antibodies GAP43 (1:5,000, Cat#: GAP43; Aves Lab); p-SAPK/JNK (T183/Y185) (1:1,000, 9251S; Cell Signaling); SAPK/JNK (1:1,000, 9252S; Cell Signaling); p44/42 MAPK (Erk1/2) (1:1,000, 9102S; Cell Signaling); pERK (1:1,000, sc-7383; Santa Cruz); p-Akt (S473) (1:1,000, 4060S; Cell Signaling); Akt, (9272S; Cell Signaling); and tubulin-HRP (1:50,000, AC030; ABclonal), diluted in 1% BSA/TBST buffer, at 4°C overnight.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Expressing

Journal: Life Science Alliance

Article Title: Nrg1 intracellular signaling regulates the development of interhemispheric callosal axons in mice

doi: 10.26508/lsa.202302250

Figure Lengend Snippet: (A) Representative images of Western blot (WB) analysis showing the expression levels of total and phosphorylated AKT, JNK, and ERK proteins in control (Nrg1 flox/flox ) and Nrg1-deficient (Nes-Cre; Nrg1 flox/flox ) neurons at DIV4. Tubulin levels are shown as a loading control. (B) Quantification of total and phosphorylated AKT, JNK, and ERK protein levels from the WB analysis, normalized to tubulin. Ctrl: n = 2; Nrg1 KO: n = 3, littermates. Unpaired t tests, in all cases, P > 0.29. Data are presented as the mean ± SEM. (C) Representative images and schematic drawings of control, Nrg1-deficient (Nrg1 KO), and Nrg1 KO neurons expressing GAP43, including the drawings used for Sholl analysis of the dendrites. Cells were fixed at DIV4. Scale bar, 50 μm. (D) Graphs showing the quantification of dendrite length in control, Nrg1 KO, and Nrg1 KO neurons expressing GAP43, based on Sholl analysis. Data are presented as the mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparisons test was used to compare Ctrl versus Nrg1 KO– and GAP43-expressing Nrg1 KO neurons. Ctrl: n = 45; Nrg1 KO: n = 44; Nrg1 KO+GAP43: n = 37, individual neurons from two different experiments. ns, not significant. (E) Quantification of axonal length in control, Nrg1 KO, and Nrg1 KO neurons expressing GAP43, expressed in microns. Ctrl, n = 104: Nrg1 KO: n = 105; GAP43: n = 78, neurons from two independent experiments. Data are presented as the mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparisons test, ** P < 0.001, *** P < 0.001.

Article Snippet: Proteins from gels were transferred onto PVDF membranes (03010040001; Roche), and they were incubated with the primary antibodies GAP43 (1:5,000, Cat#: GAP43; Aves Lab); p-SAPK/JNK (T183/Y185) (1:1,000, 9251S; Cell Signaling); SAPK/JNK (1:1,000, 9252S; Cell Signaling); p44/42 MAPK (Erk1/2) (1:1,000, 9102S; Cell Signaling); pERK (1:1,000, sc-7383; Santa Cruz); p-Akt (S473) (1:1,000, 4060S; Cell Signaling); Akt, (9272S; Cell Signaling); and tubulin-HRP (1:50,000, AC030; ABclonal), diluted in 1% BSA/TBST buffer, at 4°C overnight.

Techniques: Western Blot, Expressing, Control

Journal: Pathophysiology

Article Title: RhoG-Binding Domain of Elmo1 Ameliorates Excessive Process Elongation Induced by Autism Spectrum Disorder-Associated Sema5A

doi: 10.3390/pathophysiology30040040

Figure Lengend Snippet: Key materials used in this study.

Article Snippet: Ant-growth-associated protein 43 (GAP43) , Santa Cruz Biotechnology (Santa Cruz, CA, USA) , sc-17790 , J0920 , Immunoblotting (IB), 1:5000.

Techniques: Western Blot, Immunofluorescence, Membrane, Neutralization, Concentration Assay, Recombinant, Generated, Synthesized, Binding Assay

Journal: Pathophysiology

Article Title: RhoG-Binding Domain of Elmo1 Ameliorates Excessive Process Elongation Induced by Autism Spectrum Disorder-Associated Sema5A

doi: 10.3390/pathophysiology30040040

Figure Lengend Snippet: Mutated Sama5A excessively increases neuronal differentiation marker expression. ( A , B ) The lysates of cells following the induction of differentiation were immunoblotted with an antibody against a neuron differentiation marker protein (GAP43 or Tau), His-tag, or the internal marker actin protein. Their immunoreactive band intensities are statistically depicted (* p < 0.05, ** p < 0.01; n = 3 blots). An anti-Tau antibody recognizes approximately 50 kDa of many Tau protein variants. Other immunoreactive bands can be non-specific ones.

Article Snippet: Ant-growth-associated protein 43 (GAP43) , Santa Cruz Biotechnology (Santa Cruz, CA, USA) , sc-17790 , J0920 , Immunoblotting (IB), 1:5000.

Techniques: Marker, Expressing

Journal: Pathophysiology

Article Title: RhoG-Binding Domain of Elmo1 Ameliorates Excessive Process Elongation Induced by Autism Spectrum Disorder-Associated Sema5A

doi: 10.3390/pathophysiology30040040

Figure Lengend Snippet: RBD recovers excessive neuronal differentiation marker expression induced by Sama5A harboring the R676C mutation. ( A , B ) The lysates of cells following the induction of differentiation were immunoblotted with an antibody against GAP43, Tau, His-tag, GFP-tag, or actin protein. Their immunoreactive band intensities are statistically depicted (** p < 0.01; n = 3 blots). An anti-Tau antibody recognizes approximately 50 kDa of many Tau protein variants. Other immunoreactive bands can be non-specific ones.

Article Snippet: Ant-growth-associated protein 43 (GAP43) , Santa Cruz Biotechnology (Santa Cruz, CA, USA) , sc-17790 , J0920 , Immunoblotting (IB), 1:5000.

Techniques: Marker, Expressing, Mutagenesis