control shrna plasmid  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Occludin Is Essential to Maintain Normal Alveolar Barrier Integrity and Its Protective Role During ARDS Progression

doi: 10.3390/ijms252111595

Figure Lengend Snippet: Occludin expression is decreased after electroporation-mediated delivery of occludin shRNA plasmids. Control or occludin shRNA plasmids were delivered to the lungs of mice and, four days later, protein levels were determined in whole lung by Western blot (n = 4). Densitometry was performed and mean ± standard deviation is shown along with the individual animal data points. *, p ≤ 0.04 as determined by unpaired t -test.

Article Snippet: Four distinct RFP-expressing occludin shRNA plasmids (catalog # TF501526) and a scrambled shRNA plasmid (catalog # TR30015) were purchased from Origene (Rockville, MD, USA).

Techniques: Expressing, Electroporation, shRNA, Control, Western Blot, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Occludin Is Essential to Maintain Normal Alveolar Barrier Integrity and Its Protective Role During ARDS Progression

doi: 10.3390/ijms252111595

Figure Lengend Snippet: Electroporation-mediated gene delivery of occludin shRNAs can accelerate LPS-induced lung injury. A combination of four distinct occludin shRNA plasmids (160 μg total) in 50 μL was delivered to the lungs by electroporation at 200 V/cm using 8 pulses of 10 msec in duration. Four days after gene delivery, LPS (5 mg/kg) was intratracheally administered to mice, and two days later, lungs were removed for gravimetric analysis. ( A ) Wet-to-dry ratios are shown as mean ± SEM (n = 5). Statistical analysis was by one way ANOVA followed by post-hoc analysis using Tukey’s test, ** p < 0.01. ( B ) Hematoxylin and eosin staining were used to compare the histologic features. Scale bar: 100 µm.

Article Snippet: Four distinct RFP-expressing occludin shRNA plasmids (catalog # TF501526) and a scrambled shRNA plasmid (catalog # TR30015) were purchased from Origene (Rockville, MD, USA).

Techniques: Electroporation, shRNA, Staining

Journal: International Journal of Molecular Sciences

Article Title: Occludin Is Essential to Maintain Normal Alveolar Barrier Integrity and Its Protective Role During ARDS Progression

doi: 10.3390/ijms252111595

Figure Lengend Snippet: Gene delivery of occludin shRNA plasmids enhances LPS-induced inflammation. A 160 μg combination of 4 distinct occludin shRNA plasmids in 50 μL was delivered to the lungs by electroporation at 200 V/cm using 8 pulses of 10 msec in duration. Four days after gene delivery, LPS (5 mg/kg) was intratracheally administered to mice, and two days later, BAL fluid was collected and assessed ( A ) by cellularity of BAL fluid and ( B ) total protein levels in BAL (n = 5–6). Statistical analysis was by one way ANOVA followed by post-hoc analysis using Tukey’s test. # p < 0.05 and ** p < 0.01.

Article Snippet: Four distinct RFP-expressing occludin shRNA plasmids (catalog # TF501526) and a scrambled shRNA plasmid (catalog # TR30015) were purchased from Origene (Rockville, MD, USA).

Techniques: shRNA, Electroporation

Journal: International Journal of Molecular Sciences

Article Title: Occludin Is Essential to Maintain Normal Alveolar Barrier Integrity and Its Protective Role During ARDS Progression

doi: 10.3390/ijms252111595

Figure Lengend Snippet: Occludin is needed for alveolar barrier function in vitro. ( A ) Alveolar epithelial type II (ATII) cells transfected with siRNA-occludin or scrambled control were plated in transwell plates. TEER was measured as indicated, post transfection. Statistics were performed using two-way ANOVA with Tukey’s multiple comparisons test. n = 6, ** p < 0.01, **** p < 0.0001. ( B ) ATII cells transfected with pcDNA3 or occludin plasmid were plated in transwell plates. TEER was measured 48 h after transfection. Statistics were performed using Student’s t test (n = 6, *** p < 0.001). ( C ) Knockdown of occludin reduces claudin18 localized at tight junctions in ATII cells. ATII cells isolated from rats were electroporated with scramble or occludin shRNA. Then, 72 h later, cells were stained for occludin (red) and claudin18 (green). Nuclei are stained with DAPI (blue). Scale bar: 40 µm. ( D ) Overexpression of occludin induces claudin18 localized at tight junctions in ATII cells. ATII cells isolated from rats were electroporated with empty plasmids or plasmids expressing occludin. Then, 48 h later, cells were stained for occludin (red) and claudin18 (green). Nuclei are stained with DAPI (blue). Scale bar: 40 µm.

Article Snippet: Four distinct RFP-expressing occludin shRNA plasmids (catalog # TF501526) and a scrambled shRNA plasmid (catalog # TR30015) were purchased from Origene (Rockville, MD, USA).

Techniques: In Vitro, Transfection, Control, Plasmid Preparation, Knockdown, Isolation, shRNA, Staining, Over Expression, Expressing

Journal: STAR Protocols

Article Title: Protocol for evaluation of tumor-derived exosome-induced cancer cell metastasis in a mouse model

doi: 10.1016/j.xpro.2023.102444

Figure Lengend Snippet:

Article Snippet: Scrambled shRNA control plasmid , OriGene , Cat# TR30021.

Techniques: Recombinant, BIA-KA, Western Blot, shRNA, Control, Plasmid Preparation, Software, Modification