anti tex14 rabbit polyclonal antibody  (Proteintech)

Journal: bioRxiv

Article Title: Branched germline cysts and female-specific cyst fragmentation facilitate oocyte determination in mice

doi: 10.1101/2022.11.17.516957

Figure Lengend Snippet: (A) Time course of oocyte and gonocyte differentiation in mouse fetal gonads. A single low dose of Tamoxifen (Tmx) was injected to CAG-cre/Esr1: R26R-EYFP mice at E10.5 to label one primordial germ cell (PGC) per gonad on average. Fetal gonads were collected at E12.5, E14.5, E17.5 and P0 for cyst structure analysis. (B) Z-stack confocal images of lineage-labeled germ cell clones (left) and the cyst structure of the clone (right) showing the geometry of YFP-positive germ cells (green circle) and TEX14(or RacGAP)-positive bridges (red line). The cyst outlined by the blue dotted line was defined as sister germ cells that are connected by intercellular bridges. Each single germ cell was counted as a cyst. The clone outlined by the magenta dotted line was defined as sister germ cells derived from a single PGC labelled by lineage marker YFP. (C) Z-stack confocal images of lineage-labeled cysts showing an example of a branching cell (C (a)) and an example of unconnected bridges (C(b)). (D-E) Size of germ cell clones in fetal ovaries (D) and testes (E). (F-G) Number of germline cysts in a clone in fetal ovaries (F) and testes (G). (H-I) Size of germline cysts in fetal ovaries (H) and testes (I). (J-K) Percentage of unconnected bridges in fetal ovaries (J) and testes (K). (L-S) Percentage of the cysts profiled by the size of cysts (left Y axis) and percentage of the cysts that are in the branched structure (right Y axis) in fetal ovaries at E12.5 (L), E14.5 (M), E17.5 (N), P0 (O), and fetal testes at E12.5 (P), E14.5 (Q), E17.5 (R), P0 (S). (T-AA) Germline cysts profiled by cyst size and the number of branching germ cells contained in fetal ovaries at E12.5 (T), E14.5 (U), E17.5 (V), P0 (W), and fetal testes at E12.5 (X), E14.5 (Y), E17.5 (Z), P0 (AA). Scale bar=10 μm. The bar in the graph represents the average value. Data are presented as mean ± SD.

Article Snippet: The following primary antibodies and the dilutions (in parentheses) were used in this study: anti-GFP chicken polyclonal antibody (Aves Labs, GFP-1020, 1:1000), anti-TEX14 rabbit polyclonal antibody (ProteinTech, 18351-1-AP, 1:200), anti-RacGAP1 monoclonal antibody (Santa Cruz biotechnology, sc-271110, 1:200), anti-GM130 (BD biosciences, 610822, 1:200), anti-gamma tubulin (abcam, ab179503, 1:200), anti-E-cadherin rat monoclonal antibody (Takara, M108, 1:500), anti-GFRα1 goat polyclonal antibody (R&D Systems, AF560, 1:200), anti-PLZF mouse monoclonal antibody (EMD Millipore, OP128, 1:100), anti-UTF1 rabbit polyclonal antibody (abcam, ab24273, 1:500), anti-KIT rat monoclonal antibody (BD Biosciences, 553352, 1:200), anti-DDX4 rabbit polyclonal antibody (abcam, ab13840, 1:400) and anti-SCP3 rabbit polyclonal antibody (abcam, ab15092, 1:200).

Techniques: Injection, Labeling, Clone Assay, Derivative Assay, Marker

Journal: bioRxiv

Article Title: Branched germline cysts and female-specific cyst fragmentation facilitate oocyte determination in mice

doi: 10.1101/2022.11.17.516957

Figure Lengend Snippet: (A) Diagram showing the strategy for lineage-labeling individual Pax7-positive spermatogonia stem cells (SSCs) in the adult testis. Testes were collected on day1, 10 and 20 after Tamoxifen (Tmx) injection. (B) Confocal image showing a lineage-labelled YFP-positive undifferentiated spermatogonia expressing E-cadherin (red) one day after Tmx injection. (C) Composition of lineage-labeled spermatogonia containing different numbers of germ cells one day after Tmx injection. As: single SSC; Apr: two connected SSCs; Aal-3: three aligned SSC; Aal-4: four aligned SSCs. (D-F) Confocal images showing lineage-labeled germ cell clones that are E-cadherin positive (D), KIT positive (E) or SCP-3 positive (F) observed in the testes 10 days after Tmx injection. (G) Confocal image showing meiotic germ cells labeled by a SPC3 antibody and intercellular bridges labeled by a TEX14 antibody. (H-K) Structure of germline cysts profiled by the percentage of germ cells connected by 0, 1, 2, 3, and 4 bridges in E14.5 testes (H), E-cadherin-positive undifferentiated spermatogonia (I), KIT-positive differentiating spermatogonia (J) and spermatocytes (K). 1128 cells from 25 clones, 722 from 118 clones, 798 from 22 clones, and 640 cells were analyzed in (I), (J), (K), and (L) respectability. Scale bar=10 μm.

Article Snippet: The following primary antibodies and the dilutions (in parentheses) were used in this study: anti-GFP chicken polyclonal antibody (Aves Labs, GFP-1020, 1:1000), anti-TEX14 rabbit polyclonal antibody (ProteinTech, 18351-1-AP, 1:200), anti-RacGAP1 monoclonal antibody (Santa Cruz biotechnology, sc-271110, 1:200), anti-GM130 (BD biosciences, 610822, 1:200), anti-gamma tubulin (abcam, ab179503, 1:200), anti-E-cadherin rat monoclonal antibody (Takara, M108, 1:500), anti-GFRα1 goat polyclonal antibody (R&D Systems, AF560, 1:200), anti-PLZF mouse monoclonal antibody (EMD Millipore, OP128, 1:100), anti-UTF1 rabbit polyclonal antibody (abcam, ab24273, 1:500), anti-KIT rat monoclonal antibody (BD Biosciences, 553352, 1:200), anti-DDX4 rabbit polyclonal antibody (abcam, ab13840, 1:400) and anti-SCP3 rabbit polyclonal antibody (abcam, ab15092, 1:200).

Techniques: Labeling, Injection, Expressing, Clone Assay

Journal: bioRxiv

Article Title: Branched germline cysts and female-specific cyst fragmentation facilitate oocyte determination in mice

doi: 10.1101/2022.11.17.516957

Figure Lengend Snippet: (A-C) The volume of single germ cells and germ cells in cysts in E14.5, E17.5, and P0 germ cell clones. The bar in the graph represents the average value; the number above the bar represents the average volume of the germ cells; the number in the parentheses represents the number of measured germ cells. (D) Comparison of the germ cell volume between the germ cells with a B-body and without a B-body in P0 clones. Data in the graph are presented as mean ± SD and a t-test was used for statistical analysis. (E) Z-stack confocal image showing a lineage-labeled germ cell clone. Germ cells were revealed by YFP antibody staining, bridges were stained with a TEX14 antibody, and Golgi complexes were stained with a GM130 antibody. (F) Diagram of the germ cell clone in (E) showing the geometry of the germ cells (green circles) and the bridges (red lines) connected with them. The numbers in the germ cells represent the normalized value of the content of Golgi complexes in each germ cell. (G-I) Cyst germ cells were profiled based on the number of bridges connected with them and the amount of Golgi complexes each germ cell contained in 4-cell cysts (G), 3-cell cysts (H) and 2-cell cysts (I). (J-L) Confocal images showing a lineage-labeled germ cell clone in the P0 testis. The testis was stained with a GFP antibody for cyst germ cells, a TEX14 antibody for intercellular bridges and an antibody for spermatogonia stem cell marker GFRα1. Arrowheads indicate lineage-labeled germ cells with negative GFRα1 staining. (M-N) Percentage of cysts (M) and clones (N) containing the germ cells with only GFRα1 positive germ cells (red box), only GFRα1 negative germ cells (white box) and both GFRα1 positive and negative germ cells (stripped box). Scale bar=10 μm.

Article Snippet: The following primary antibodies and the dilutions (in parentheses) were used in this study: anti-GFP chicken polyclonal antibody (Aves Labs, GFP-1020, 1:1000), anti-TEX14 rabbit polyclonal antibody (ProteinTech, 18351-1-AP, 1:200), anti-RacGAP1 monoclonal antibody (Santa Cruz biotechnology, sc-271110, 1:200), anti-GM130 (BD biosciences, 610822, 1:200), anti-gamma tubulin (abcam, ab179503, 1:200), anti-E-cadherin rat monoclonal antibody (Takara, M108, 1:500), anti-GFRα1 goat polyclonal antibody (R&D Systems, AF560, 1:200), anti-PLZF mouse monoclonal antibody (EMD Millipore, OP128, 1:100), anti-UTF1 rabbit polyclonal antibody (abcam, ab24273, 1:500), anti-KIT rat monoclonal antibody (BD Biosciences, 553352, 1:200), anti-DDX4 rabbit polyclonal antibody (abcam, ab13840, 1:400) and anti-SCP3 rabbit polyclonal antibody (abcam, ab15092, 1:200).

Techniques: Clone Assay, Comparison, Labeling, Staining, Marker

Journal: Developmental biology

Article Title: Deletion of MgcRacGAP in the male germ cells impairs spermatogenesis and causes male sterility in the mouse.

doi: 10.1016/j.ydbio.2013.12.006

Figure Lengend Snippet: Fig. 5. A. Transmission electronic microscopic view of an intercellular bridge (red arrow) from a 10 dpp normal testis and of a multinuclear cell from a 10 dpp knock out (KO) testis. No bridges could be observed in the KO testes. Scale bar: 1 μm. B. TEX14 labelling of control (Ct) and knock out (KO) testes at 8 dpp. Red: TEX14, blue: DAPI staining of the nuclei. White arrows indicate the intercellular bridges labelled with Tex14 antibody in the control testis. A faint punctate and non-specific staining is also detected in the nucleus of both control and mutant testes. C. Western blot analysis of TEX14 protein amount in control (Ct) and knock out (KO) testes.

Article Snippet: The following antibodies and dilutions were used: chicken polyclonal anti-MgcRacGAP (described in Toure et al. (2001); 1/1000), guinea pig anti-Vimentin antibody (Progen, 1/1000), rabbit polyclonal anti-Tex14 antibody (Abcam, 1/200), rabbit anti-LC3B antibody (L7543 Sigma, 1/1000) mouse anti-Atg12 antibody (#2011 Cell Signaling, 1/1000), mouse monoclonal anti-βtubulin antibody (Sigma, 1/1000), rabbit polyclonal anti-mouse Ig coupled to Horse Radish Peroxydase (HRP) (Dako, 1/1000), swine polyclonal antirabbit Ig coupled to HRP (Dako, 1/500), rabbit polyclonal antiguinea pig Ig coupled to HRP (Dako, 1/1000) and rabbit antichicken Ig coupled to HRP (Pierce, 1/30,000).

Techniques: Transmission Assay, Knock-Out, Control, Staining, Mutagenesis, Western Blot