Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: The primers used in present study.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques:

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: RasGRF1 expression in hippocampus dramatically decreased after chronic cerebral hypoperfusion. ( A ) The RasGRF1 expression, the total and phosphorylated ERK expression were assayed by Western blot. β-actin was as inner control for samples loading normalization; (Con ( n = 3), 2VO ( n = 3)). ( B ) The RasGRF1 relative expression level was calculated and analyzed statistically. ( C ) The p-ERK relative expression level was calculated and analyzed statistically. Relative intensity of p-ERK level is calculated by p-ERK/total ERK. Total ERK was as inner control for samples loading normalization. ( D ) RasGRF1 distribution in brain was observed with immunohistochemical staining; (Con ( n = 1), 2VO ( n = 1)). ( E ) In hippocampal subregions and cortex, the RasGRF1-positive cells were counted and analyzed. ( F ) The neurons number was observed with Nissl staining; (Con ( n = 1), 2VO ( n = 1)). ( G ) The Nissl-stained positive cells were counted and analyzed. Compared with Control, ** p < 0.01. Scale bar = 50 μm. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Expressing, Western Blot, Control, Immunohistochemical staining, Staining

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: miR-323-3p could regulate the expression of RasGRF1 through binding to the 3′UTR. The total RNAs were extracted, and RasGRF1 and the predicted bind miRNAs levels were measured by RT-PCR. ( A – G ) The correlation between miRNAs (rno-miR-137-3p, rno-miR-195-3p, rno-miR-203a-3p, rno-miR-21-3p, rno-miR-323-3p, rno-miR-375-3p, rno-miR-448-5p) and RasGRF1 levels were analyzed. ( H ) The correlation degree was displayed with heatmap. ( I ) rno-miR-323-3p, rno-miR-375-3p and rno-miR-448-5p level in hippocampus after CCH were showed. ( J ) Dual luciferase reporter assayed the binding and expression regulation of targeted RasGRF1 mRNAs 3’UTR by miRNA mimics and negative control of miRNA-323-3p, miRNA-375-3p and miRNA-448-5p. ( K ) The binding and regulation between mutated 3’UTR of RasGRF1 mRNA and miRNA-323-3p was assayed by dual luciferase reporter. ( L ) RasGRF1 level after rno-miR-323-3p mimic treatment was detected by Western blot. β-actin was as inner control for samples loading normalization. ( M ) The RasGRF1 relative expression level was calculated and analyzed statistically. Compared with miRNA-Control, ** p < 0.01. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Expressing, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Luciferase, Negative Control, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: The inhibition of miR-323-3p could up-regulate the expression of RasGRF1 in hippocampus after CCH. ( A ) The RasGRF1 expression, the total and phosphorylated ERK expression were assayed by Western blot. β-actin was as inner control for samples loading normalization. (Con ( n = 3), 2VO ( n = 3), 2VO+miRNA-323-3p antagomir ( n = 3), 2VO+ miRNA-323-3p antagomir NC ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 siRNA ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 scramble siRNA ( n = 3)). ( B ) The relative expression level of RasGRF1 was calculated and analyzed statistically. ( C ) The p-ERK relative expression level was calculated and analyzed statistically. Relative intensity of p-ERK level is calculated by p-ERK/total ERK. ** p < 0.01. The experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition, Expressing, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: Rasgrf1 upregulation by the inhibition of miR-323-3p could improve the spatial cognitive. dysfunction after CCH. MWM was employed to detect the spatial cognitive abilities. ( A ) The latency to reach the platform during the trail training was recorded and analyzed. ( B ) After 1 day-rest, the latency to reach the platform. ( C ) The crossing times around the platform, and ( D ) staying time in platform quadrant were recorded and analyzed. The Y maze was employed to further test the cognitive abilities. ( E ) The number of arms entries alteration, ( F ) percent of arms entries alteration, and ( G ) total arms entries times were recorded and analyzed. Con: sham group, n = 12; 2VO: bilateral common arteries occlusion group, n = 15; 2VO+miR-323-3p antagomir: bilateral common arteries occlusion group with miR-323-3p antagomir intra-cerebroventricular injection (ICV) injection, n = 15; 2VO+miR-323-3p NC: 2VO group with ICV injection of miR-323-3p antagomir negative control, n = 15; 2VO+miR-323-3p antagomir+RasGRF1 siRNA: 2VO group with ICV injection of miR-323-3p antagomir and RasGRF1 siRNA, n = 15; 2VO+miR-323-3p antagomir + RasGRF1 scramble: 2VO group with ICV injection of miR-323-3p antagomir and RasGRF1 siRNA scramble, n = 15. * p < 0.05, ** p < 0.01. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition, Injection, Negative Control

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: Rasgrf1 upregulation by the inhibition of miR-323-3p could improve the long-term potential impairment after CCH. ( A ) Field excitable postsynaptic potential was recorded and analyzed pre- and post- high frequency stimulation (pre-HFS recording as the baseline). (Con ( n = 3), 2VO ( n = 3), 2VO+miRNA-323-3p antagomir ( n = 3), 2VO+ miRNA-323-3p antagomir NC ( n = 3), 2VO+miRNA-323-3p antagomir+RasGRF1 siRNA ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 scramble siRNA ( n = 3)). ( B ) Relative field excitable postsynaptic potential and ( C ) population spike was analyzed over the baseline. ** p < 0.01. The experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: Rasgrf1 upregulation by the inhibition of miR-323-3p could ameliorate the neural structural plasticity impairment after CCH. ( A ) The dendritic spines were observed with Golgi staining. ( B ) The cumulative distribution of dendritic spines density was calculated; (Con ( n = 1), 2VO ( n = 1), 2VO+miRNA-323-3p antagomir ( n = 1), 2VO+ miRNA-323-3p antagomir NC ( n = 1), 2VO+miRNA-323-3p antagomir + RasGRF1 siRNA ( n = 1), 2VO+miRNA-323-3p antagomir + RasGRF1 scramble siRNA ( n = 1)). ( C ) The total dendritic spines density. ( D ) The cumulative distribution of dendritic spines density. ( E ) The mushroom dendritic spines density was counted and calculated. Scale bar = 5 μm. ** p < 0.01. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition, Staining

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: Rasgrf1 upregulation by the inhibition of miR-323-3p could improve the synapse density decrease after CCH. ( A ) The synapses were observed with uranyl acetate negative staining under transmission electron microscope; (Con ( n = 3), 2VO ( n = 3), 2VO+miRNA-323-3p antagomir ( n = 3), 2VO+miRNA-323-3p antagomir NC ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 siRNA ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 scramble siRNA ( n = 3)). ( B ) Synapses densities were calculated. ( C and D ) The cumulative and specific distribution of synapses density was calculated. Scale bar = 200 nm. ** p < 0.01. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition, Negative Staining, Transmission Assay, Microscopy

Journal: Aging (Albany NY)

Article Title: Upregulation of RasGRF1 ameliorates spatial cognitive dysfunction in mice after chronic cerebral hypoperfusion

doi: 10.18632/aging.204654

Figure Lengend Snippet: Rasgrf1 upregulation by the inhibition of miR-323-3p could improve the synapse structure deterioration after CCH. ( A ) The synapses were observed with uranyl acetate negative staining under transmission electron microscope and the synapses’ structure was observed; (Con ( n = 3), 2VO ( n = 3), 2VO+miRNA-323-3p antagomir ( n = 3), 2VO+ miRNA-323-3p antagomir NC ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 siRNA ( n = 3), 2VO+miRNA-323-3p antagomir + RasGRF1 scramble siRNA ( n = 3)). ( B and C ) The cumulative and specific distribution of synaptic PSD length. ( D ) PSD length of synapses. ( E ) Synapse area and ( F ) the relative PSD intensity of synapses were measured and calculated. Scale bar = 200 nm. ** p < 0.01. All of the experiments were repeated three times.

Article Snippet: The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry.

Techniques: Inhibition, Negative Staining, Transmission Assay, Microscopy

Journal: Animals : an Open Access Journal from MDPI

Article Title: Changes in Tight Junction Protein Expression Levels but Not Distribution in Commercial White and Brown Laying Hens Supplemented with Chondrus crispus or Ascophyllum nodosum Seaweed

doi: 10.3390/ani14050777

Figure Lengend Snippet: Tight junction regulatory protein immunoreactivity in hen jejunum. ( a ) Occludin (OCC) reactivity (red) is seen throughout the intestinal epithelium, appearing more intense in the deeper cryptal areas adjacent to the muscularis. ( b ) At higher magnification, OCC staining can be seen across the cell surface of the epithelial layer, often concentrated at the apicolateral cell surfaces. ( c ) OCC negative control. ( d ) Expression of zonula occludens-1 (ZO-1) protein in jejunal epithelium, appearing less pronounced in cryptal areas than the luminal villi. ( e ) Higher magnification shows ZO-1 reactivity is most pronounced at the apico-lateral cell surface. ( f ) ZO-1 negative control. v—villus; c—crypt; m—muscularis.

Article Snippet: Sections were incubated for 45 min with 1.25 mg/mL primary rabbit polyclonal antibody against occludin (Cat.# 711500, Fisher Scientific), 1.25 mg/mL rabbit polyclonal antibody ZO-1 (Cat.#617300, Fisher Scientific) or 1.25 mg/mL control rabbit IgG (Cat #026102, Fisher Scientific).

Techniques: Staining, Negative Control, Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Changes in Tight Junction Protein Expression Levels but Not Distribution in Commercial White and Brown Laying Hens Supplemented with Chondrus crispus or Ascophyllum nodosum Seaweed

doi: 10.3390/ani14050777

Figure Lengend Snippet: Western blot results of occludin (OCC) and zonula occludens-1 (ZO-1) protein expression in laying hen jejunum. Graphed results are least squares means ± standard error following Tukey’s means comparisons of square-root transformed relative band intensities. ( a ) Strain and seaweed supplement effects; within effect grouping, means with the same letter were not different ( p < 0.05). ( b ) Strain x seaweed interaction effect on ZO-1 expression. Co—control diet; CC— Chondrus crispus supplement; AN— Ascophyllum nodosum supplement (See for original western blot information of ).

Article Snippet: Sections were incubated for 45 min with 1.25 mg/mL primary rabbit polyclonal antibody against occludin (Cat.# 711500, Fisher Scientific), 1.25 mg/mL rabbit polyclonal antibody ZO-1 (Cat.#617300, Fisher Scientific) or 1.25 mg/mL control rabbit IgG (Cat #026102, Fisher Scientific).

Techniques: Western Blot, Expressing, Transformation Assay, Control