cox1 shrna plasmid for a mouse sc-35097-sh  (Santa Cruz Biotechnology)

Journal: Andrology

Article Title: Differential roles of cyclooxygenase enzymes in the regulation of murine juvenile undifferentiated spermatogonia.

doi: 10.1111/andr.13537

Figure Lengend Snippet: FIGURE 3 Characterization of C18-4Cox1-KD1 cells and KD-associated morphological and gene changes. (A) Expression of cyclooxygenase (Cox)1 on populations of transfected cells with varying concentrations of shRNA and lipofectamine 3000 showing that knockdown “B” had the most downregulated expression of Cox1. (B) Cox1 protein expression on immunoblot, comparing wildtype cells (WT), scrambled cells (Scrb), knockdown “B” clone, and an isolated clonal population of C18-4Cox1-KD1. C18-4Cox1-KD1 Cox1 protein expression validated against the total protein. (C) Gene expression of Cox1 in C18-4Cox1-KD1 and Scrambled controls, (D) Immunofluorescence staining of Cox1 in C18-4Cox1-KD1 and Scrambled controls. Green: Cox1, red: alpha-tubulin, blue: DAPI. Scale in µm. (E) Brightfield visualization illustrating morphological differences between C18-4Cox1-KD1 and Scrambled controls. (F) Differences in gene expression between C18-4Cox1-KD1 and Scrambled controls of Jam-1, Mmp2, Stra8, and Kit. Results are presented as fold change of controls. mRNA expression data were normalized to Glyceraldehyde 3-phosphate dehydrogenase (Gapdh); N = 3 independent experiments conducted in triplicates. Significant difference relative to controls with t-test: * (p ≤0.05), ** (p < 0.01), *** (p < 0.001).

Article Snippet: Cox1 shRNA plasmid for a mouse (sc-35097-SH) and control shRNA plasmid (sc-108060) were purchased from Santa Cruz Biotech.

Techniques: Expressing, Transfection, shRNA, Knockdown, Western Blot, Isolation, Gene Expression, Immunofluorescence, Staining

Journal: Andrology

Article Title: Differential roles of cyclooxygenase enzymes in the regulation of murine juvenile undifferentiated spermatogonia.

doi: 10.1111/andr.13537

Figure Lengend Snippet: FIGURE 5 Activation of Notch3 signaling pathway in C18-4Cox1-KD1. (A) Immunofluorescence (IF) images of Notch3 expression in C18-4Cox1-KD1 and Scrambled controls. Green: Notch3, red: alpha-tubulin, blue: DAPI. Scale in µm. (B) Immunoblots of Notch3, Hes1, cyclooxygenase (Cox)1, and Cox2 expression in C18-4Cox1-KD1 and Scrambled controls are cropped at indicated bands, and protein expression, indicated by values under the bands, is normalized to alpha-tubulin. (C) Quantification of Notch3, Hes1, Cox1, and Cox2 protein expression on western blot normalized against the total protein. Results are presented as fold change of controls. (D) IF images of Hes1 expression in C18-4Cox1-KD1 and Scrambled controls. Green: Hes1, red: alpha-tubulin, blue: DAPI. Hes1-positive cells were counted and the data show the percentage of Hes-positive cells as fold of control samples. N = 3 independent experiments. Significant difference relative to controls with t-test: * (p ≤0.05), ** (p < 0.01), *** (p < 0.001).

Article Snippet: Cox1 shRNA plasmid for a mouse (sc-35097-SH) and control shRNA plasmid (sc-108060) were purchased from Santa Cruz Biotech.

Techniques: Activation Assay, Immunofluorescence, Expressing, Western Blot, Control