mouse anti par3  (Santa Cruz Biotechnology)

Journal: Development (Cambridge, England)

Article Title: MMP14 is required for delamination of chick neural crest cells independently of its catalytic activity

doi: 10.1242/dev.183954

Figure Lengend Snippet: Destabilizing apicobasal polarity is sufficient to rescue siRNA-MMP14. (A) Immunostaining for snail 2 (gray) after electroporation of siRNA-MMP14 or co-electroporation with siRNA-MMP14 and Par3. Bracket indicates accumulation of neural crest cells. (B) Plot of neural crest area (snail 2) with siRNA-MMP14 (nembryos=14, nsections=361), siRNA-MMP14 with Par3 (nembryos=6, nsections=166) and siRNA-MMP14 with Par3MO (nembryos=7, nsections=38) from two independent experiments. ANOVA and uncorrected Fisher's LSD; ***P=0.0007, ****P<0.0001. Scale bars: 25 µm. Par3MO efficiency/specificity could not be checked (see main text and Materials and Methods for details).

Article Snippet: We attempted to use rabbit anti-mouse Par3 and mouse anti-human Par3 (Millipore, 07-330 and 8E8) but none of them produced staining that was specific for chicken Par3.

Techniques: Immunostaining, Electroporation

Journal: Development (Cambridge, England)

Article Title: MMP14 is required for delamination of chick neural crest cells independently of its catalytic activity

doi: 10.1242/dev.183954

Figure Lengend Snippet: Destabilizing apicobasal polarity is sufficient to rescue siRNA-MMP14. (A) Immunostaining for snail 2 (gray) after electroporation of siRNA-MMP14 or co-electroporation with siRNA-MMP14 and Par3. Bracket indicates accumulation of neural crest cells. (B) Plot of neural crest area (snail 2) with siRNA-MMP14 (nembryos=14, nsections=361), siRNA-MMP14 with Par3 (nembryos=6, nsections=166) and siRNA-MMP14 with Par3MO (nembryos=7, nsections=38) from two independent experiments. ANOVA and uncorrected Fisher's LSD; ***P=0.0007, ****P<0.0001. Scale bars: 25 µm. Par3MO efficiency/specificity could not be checked (see main text and Materials and Methods for details).

Article Snippet: We attempted to use rabbit anti-mouse Par3 and mouse anti-human Par3 (Millipore, 07-330 and 8E8) but none of them produced staining that was specific for chicken Par3.

Techniques: Immunostaining, Electroporation

Journal: eLife

Article Title: A mitotic kinase scaffold depleted in testicular seminomas impacts spindle orientation in germ line stem cells

doi: 10.7554/eLife.09384

Figure Lengend Snippet: ( A ) Immunoblot analysis of tissue lysates from resected seminomas (lanes 2, 4, and 6) and normal adjacent tissue (lanes 1, 3, and 5). Proteins were identified using antibodies against (top) Gravin, (upper-mid) Aurora A, (lower-mid) Plk1, and (bottom) GAPDH loading control. ( B ) Quantification of immunoblot data ( A ) by densitometry (n = 3 ± SEM). ( C , D ) Representative testis sections from ( C ) a 30-year-old individual and ( D ) a 26-year-old seminoma patient. Immunofluorescent staining shows Gravin (green), p-H3B (red), and DNA (DAPI, blue). Scale bar, 40 μm. ( E , F ) Magnified insets from C and D are included. Scale bar, 40 μm. ( G ) Gravin signal intensity per mitotic cell was quantified from normal and seminoma sections of testis (p-H3B positive, n-values are indicated, ***p < 0.001). The number of cells used in each analysis is indicated. ( H ) The mitotic index was calculated for (normal; n = 4) and (seminoma; n = 6) tissue sections by determining the percentage of pH3B-positive cells. (*p < 0.05). ( I , J ) Related experiments were conducted on testis sections from 7-week-old wild-type ( I ), and Gravin knockout ( J ) mice. Immunostaining with antibodies against Par3 (green), p-H3B (red), and DAPI (blue) is presented. Scale bar, 40 μm. ( K ) Calculation of the mitotic index in testis sections from wild-type (gray) and Gravin knockout (orange) mice. The number of tissue sections measured is indicated below each column (*p < 0.05). ( L ) TUNEL staining was used to monitor apoptosis in seminiferous tubule sections from wild-type (gray) and Gravin knockout (orange) mice. Data are presented as TUNEL-positive cells per seminiferous tubule. The number of sections is depicted below each column. (**p = 0.01). DOI: http://dx.doi.org/10.7554/eLife.09384.003

Article Snippet: The following antibodies were used: mouse α-tubulin, FITC-conjugated α-tubulin (Sigma, St. Louis, MO, United States), rabbit anti-pericentrin [M8; ( )], goat anti-Aurora A (Sigma), rabbit anti-p-Aurora A T288 (Cell Signaling Technology, Danvers, MA, United States), mouse anti-Aurora B (Abcam, Cambridge, MA, United States), human anti-CREST (Antibodies Incorporated, 15–234), rabbit anti-cenexin (Protein Tech Group, Chicago, IL, United States), rabbit p-Gravin T766A , mouse Gravin (Sigma; clone JP74), mouse anti-GAPDH (Sigma; GAPDH71.1), mouse anti-phospho-S10 Histone H3 (abcam; ab14955), rabbit anti-Plk1 (Cell Signaling Technology), mouse anti-Plk1 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, United States), mouse anti-Plk1 (Millipore; 35–206, Billerica, MA, United States), rabbit anti p-Plk1T210 (Cell Signaling Technology), mouse anti-Flag and Flag-HRP (Sigma), mouse anti-Par3 (Sigma), rabbit anti-Oct3/4 (Santa Cruz Biotechnology, Inc.), mouse anti-centrin (clone 20H5, EMD Millipore), mouse anti-acetylated tubulin (Sigma), mouse anti-centrobin (Abcam).

Techniques: Western Blot, Staining, Knock-Out, Immunostaining, TUNEL Assay