control open reading frame  (OriGene)

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression improved erectile function in CNI mice. (A) Schematic of the experimental procedure. (B) Representative ICP responses of sham (7 days) and CNI (7 days) mice stimulated at 10 days after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). (C, D) Ratios of mean maximum and total ICPs (area under the curve) to MSBP were calculated for each group and results are presented as mean±standard error of mean (n=8). *** p<0.001. CNI, cavernous nerve injury; ICP, intracavernous pressure; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; MSBP, mean systolic blood pressure; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, Injection, Negative Control, Saline

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression increased cavernous endothelial and pericyte numbers in CNI mice. (A) Immunofluorescence staining for CD31 (an endothelial cell marker, green) and NG2 (a pericyte marker, red) in cavernous tissues from sham, CNI mice after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). Nuclei were labeled with DAPI (blue). Scale bar=100 µm. (B, C) Quantification of CD31 and NG2 immunopositive areas in cavernous tissue was performed using ImageJ software relative to the sham control. Results are presented as mean±standard error of mean (n=7). *** p<0.001. The relative ratio in the Sham group was defined as 1. CNI, cavernous nerve injury; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; DAPI, 4,6-diamidino-2-phenylindole; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, Immunofluorescence, Staining, Marker, Injection, Negative Control, Labeling, Software, Control, Saline

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression induced nerve regeneration in CNI mice. (A) Immunofluorescence staining for nNOS (green) and NF (red) in DNB from sham, CNI mice after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). Nuclei were labeled with DAPI (blue). Scale bar=25 µm. (B, C) Quantification of nNOS and NF immunopositive areas in cavernous tissue was performed using ImageJ software, and results are presented as mean±standard error of mean (SEM) (n=7). *** p<0.001. (D) Representative western blots for Ago2, nNOS, BDNF, and NT-3 in cavernous tissues of mice in the groups mentioned above. β-Actin was used as the internal control. (E-H) Normalized band intensity values were quantified using ImageJ software relative to the sham control. Results are presented as mean±SEM (n=4). * p<0.05; ** p<0.01; *** p<0.001. The relative ratio in the Sham group was defined as 1. CNI, cavernous nerve injury; nNOS, neuronal nitric oxide synthase; NF, neurofilament; DNB, dorsal nerve bundles; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; DAPI, 4,6-diamidino-2-phenylindole; BDNF, brain-derived neurotrophic factor; NT-3, neurotrophin-3; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, Immunofluorescence, Staining, Injection, Negative Control, Labeling, Software, Western Blot, Control, Saline, Derivative Assay

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression reduced apoptosis in CNI mice. (A) TUNEL assay of cavernous tissues from sham, CNI mice after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). Nuclei were labeled with DAPI (blue). Scale bar=100 µm. (B) Representative western blots for Bcl-2, Bax, phosphor-c-Jun/c-Jun (p-c-Jun/c-Jun), and phosphor-JNK/JNK (p-JNK/JNK) in cavernous tissues of mice in the groups mentioned above. β-Actin was used as the internal control. (C) Quantification of the number of apoptotic cells in cavernous tissue was performed using ImageJ software relative to the sham control. Results are presented as mean±standard error of mean (SEM) (n=7). *** p<0.001. (D-G) Normalized band intensity values were quantified by ImageJ software and results are presented as mean±SEM (n=4). * p<0.05; ** p<0.01. The relative ratio in the Sham group was defined as 1. CNI, cavernous nerve injury; TUNEL, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; DAPI, 4,6-diamidino-2-phenylindole; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, TUNEL Assay, Injection, Negative Control, Labeling, Western Blot, Control, Software, End Labeling, Saline

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression reduced ROS production in CNI mice. (A) Immunofluorescence staining for nitrotyrosine (green, peroxynitrite) and in situ detection of hydroethidine (red, superoxide anion) in cavernous tissue from sham, CNI mice after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). Nuclei were labeled with DAPI (blue). Scale bar=100 µm. (B, C) Quantification of nitrotyrosine and hydroethidine immunopositive areas in cavernous tissue was performed using ImageJ software relative to the sham control. Results are presented as mean±standard error of mean (SEM) (n=5). *** p<0.001. (D) Representative western blots for iNOS and p47phox in cavernous tissue with the same groups as mentioned above. β-Actin was used as the internal control. (E, F) Normalized band intensity values were quantified using ImageJ software relative to the sham control. Results are presented as mean±SEM (n=4). * p<0.05; ** p<0.01; *** p<0.001. The relative ratio in the Sham group was defined as 1. CNI, cavernous nerve injury; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; DAPI, 4,6-diamidino-2-phenylindole; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, Immunofluorescence, Staining, In Situ, Injection, Negative Control, Labeling, Software, Control, Western Blot, Saline

Journal: Investigative and Clinical Urology

Article Title: Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury

doi: 10.4111/icu.20240077

Figure Lengend Snippet: Ago2 overexpression restored mitochondrial dysfunction in CNI mice. (A) Immunofluorescence staining with MitoSOX red (for mitochondria superoxide) in cavernous tissues from sham, CNI mice after an intracavernous injection with PBS (20 µL), lentiviruses containing ORF negative control particles (NC, 5×10 4 IFU/mice), or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, 5×10 4 IFU/mice). Nuclei were labeled with DAPI (blue). Scale bar=100 µm. (B) Quantification of MitoSOX immunopositive areas in cavernous tissue was performed using ImageJ software and results are presented as mean±standard error of mean (SEM) (n=7). ** p<0.01; *** p<0.001. (C) Ratios of mitochondria ATP levels in cavernous tissue were determined using an ATP assay kit and the groups mentioned above (n=5). ** p<0.01; *** p<0.001. (D) Representative western blots for MT-ND1 and MT-CO1 in the cavernous tissues of mice in the same groups. β-Actin was used as the internal control. (E, F) Normalized band intensity values were quantified using ImageJ software relative to the sham control. Results are presented as mean±SEM (n=4). * p<0.05; ** p<0.01; *** p<0.001. The relative ratio in the Sham group was defined as 1. CNI, cavernous nerve injury; PBS, phosphate-buffered saline; ORF, open reading frame; IFU, infectious units; DAPI, 4,6-diamidino-2-phenylindole; n.s., not significant.

Article Snippet: To investigate the efficacy and mechanism whereby Ago2 restores erectile function in CNI-ED mice, we injected lentiviruses containing open reading frame (ORF) negative control particles (NC, PS100092V; Origene Technologies) or lentiviruses containing an ORF mouse clone of Ago2 (Ago2 O/E, MR227026L3V; Ori-Gene Technologies) into the penises of CNI-induced ED mice at three doses (5×10 3 , 1×10 4 , or 5×10 4 IFU/mouse).

Techniques: Over Expression, Immunofluorescence, Staining, Injection, Negative Control, Labeling, Software, ATP Assay, Western Blot, Control, Saline

Journal: eLife

Article Title: Effect of SARS-CoV-2 proteins on vascular permeability

doi: 10.7554/eLife.69314

Figure Lengend Snippet: (a) Bright-field and fluorescent image of infected eGFP HUVEC, scale bar: 50 µm; ( b ) changes in barrier functions as a result of SARS-CoV-2 proteins were assessed by trans-epithelial-endothelial electrical resistance (TEER) measurement. Note the statistical differences compared to the untreated control condition, assessed by F-statistic with two-way ANOVA test, followed by the Holm–Sidak test for multiple comparisons; ( c ) color map showing a gradual decrease in TEER values compared to the untreated condition at day 3; ( d ) immunocytochemistry (ICC) for CD31 (green) and Hoechst (blue) for the three specified conditions, scale bar: 20 µm; ( e ) analysis of CD31 expression levels.

Article Snippet: Plasmids encoding the SARS-CoV-2 open reading frames proteins and eGFP control were a kind gift of Nevan Krogan (Addgene plasmid #141367–141395).

Techniques: Infection, Control, Immunocytochemistry, Expressing

Journal: eLife

Article Title: Effect of SARS-CoV-2 proteins on vascular permeability

doi: 10.7554/eLife.69314

Figure Lengend Snippet: (a) Confocal reconstructions of HUVEC stained for von Willebrand factor (VWF) (green) and Hoechst (blue) for three conditions: control (untreated), eGFP, and nsp5_c145a, scale bar: 20 µm; ( b ) analysis of VWF expression levels; ( c ) fold change of interleukin (IL)-6 in response to the different proteins.

Article Snippet: Plasmids encoding the SARS-CoV-2 open reading frames proteins and eGFP control were a kind gift of Nevan Krogan (Addgene plasmid #141367–141395).

Techniques: Staining, Control, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: miR-374a-5p inhibits non-small cell lung cancer cell proliferation and migration via targeting NCK1

doi: 10.3892/etm.2021.10375

Figure Lengend Snippet: NCK1 reverses the effects of miR-374a-5p on NSCLC cell behaviors. (A) miR-374a-5p expression in NSCLC cells with miR-374a-5p mimics or miR-con transfection. *** P<0.001 vs. the miR-con group. (B) Cell proliferation was assessed. *** P<0.001 vs. the pcDNA3.1 group, * P<0.05 vs. the pcDNA3.1 group and ### P<0.001 vs. the miR-con group. (C) cell migration (magnification, x40) was determined. ** P<0.01 vs. the pcDNA3.1 group, * P<0.05 vs. the pcDNA3.1 group and ### P<0.001 vs. the miR-con group. (D) NCK1 expression of NSCLC cells transfected with miR-374a-5p mimics, miR-con, pNCK1, pcDNA3.1 or pNCK1 and miR-374a-5p mimics is presented. *** P<0.001 and * P<0.05 vs. the pcDNA3.1 group. NCK1, NCK adaptor protein 1; pNCK1, NCK1 overexpression plasmid; pcDNA3.1, empty vector; miR-374a-5p, microRNA-374a-5p; NSCLC, non-small cell lung cancer; miR-con, negative control miR; OD, optical density.

Article Snippet: NSCLC cells were transfected with 50 nM miR-374a-5p mimics (cat. no. miR10000727-1-5), negative control miRNA (miR-con; cat. no. miR1N0000001-1-5), 4 µg pcDNA3.1 containing the open reading frame of NCK1 (pNCK1) or negative control (pcDNA3.1) purchased from RiboBio with Lipofectamine ® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) at 37˚C.

Techniques: Expressing, Transfection, Migration, Over Expression, Plasmid Preparation, Negative Control