rabbit anti foxl2  (Proteintech)

Journal: The EMBO Journal

Article Title: MCM8 interacts with DDX5 to promote R-loop resolution

doi: 10.1038/s44318-024-00134-0

Figure Lengend Snippet: ( A ) Immunostaining of DDX4 (a marker for late germ cells) and FOXL2 (a marker for granulosa cells) in the gonads of Mcm8 +/+ and Mcm8 −/− mice at E13.5, E15.5, and PD3. Scale bars: 50 μm. ( B ) Immunostaining of DDX4 and SOX9 (a marker for Sertoli cells) in the gonads of Mcm8 +/+ and Mcm8 −/− mice at E13.5 and PD3. Scale bars: 50 μm. ( C ) Representative images of alkaline phosphatase (a PGC marker) staining of whole-mount or genital ridges from Mcm8 +/+ and Mcm8 −/− embryos. Scale bars: 200 μm. ( D ) PGC counts from Mcm8 +/+ and Mcm8 −/− embryos at E8.5, E9.5, and E11.5. n = 6/6/7/7/7/11 embryos. Data information: In ( D ), data are presented as the mean ± SD, and the dots indicate individual embryos. The statistical significance of the difference was analyzed by unpaired two-tailed Student’s t test ( D ), and the P ‐values were shown. .

Article Snippet: The primary antibodies used for immunostaining were goat anti-STELLA (1:50 dilution; Novus Biologicals AF2566), goat anti-DDX4 (1:50 dilution; Novus Biologicals AF2030), rabbit anti-FOXL2 (1:250 dilution; Abcam ab246511), rabbit anti-SOX9 (1:200 dilution; Millipore AB5535), rabbit anti-cleaved PARP1 (1:100 dilution; Cell Signaling Technology 94885), rabbit anti-Ki67 (1:200 dilution; Abcam ab15580), rabbit anti-Cyclin B1 (1:200 dilution; Cell Signaling Technology 4138), rabbit anti-53BP1 (1:500 dilution; Novus Biologicals NB100-304), rabbit anti-γH2AX (1:50 dilution; Cell Signaling Technology 9718), rabbit anti-pATM (1:100 dilution; Cell Signaling Technology 5883), rabbit anti-pSer2 Pol II (1:100 dilution; Abcam Ab193468), rabbit anti-pSer5 Pol II (1:100 dilution; Abcam Ab193467), rabbit anti-5mC (1:100 dilution; Abcam Ab214727), rabbit anti-H3K9me2 (1:100 dilution; Millipore 07-441), rabbit anti-H3K27me3 (1:100 dilution; Millipore 07-449), rabbit anti-S9.6 (1:200 dilution; Kerafast Kf-Ab01137-23.0), goat anti-S9.6 (1:400 dilution; Kerafast Kf-Ab01137-24.1), mouse anti-Fibrillarin (1:500 dilution; Abcam ab4566), rabbit anti-DDX5 (1:400 dilution; Abcam ab21696), rabbit anti-DHX9 (1:400 dilution; Abcam ab26271), and mouse anti-HA (1:200 dilution; Cell Signaling Technology 2367).

Techniques: Immunostaining, Marker, Staining, Two Tailed Test

Journal: eLife

Article Title: DMRT1 is a testis-determining gene in rabbits and is also essential for female fertility

doi: 10.7554/elife.89284

Figure Lengend Snippet: Figure 3. Somatic markers location and expression during ovarian differentiation. (A) Immunostaining of DMRT1 on XX control ovaries from 14 to 18 dpc. (B) Quantitative RT-PCR (RT-qPCR) analyses of RSPO1, DMRT1, and FOXL2 expression from 16 to 20 dpc in control gonads of both sexes. The error bars correspond to the standard error of the mean (n=3-5) (C) RSPO1 in situ hybridization (RNAscope technology), immunostaining of DMRT1 and FOXL2 on 20 dpc control ovaries. Scale bar = 50 µm.

Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Biological sample (Oryctolagus cuniniculus) Gonads Hypharm NZ1777 New Zealand rabbits Commercial kit RNAscope kit ACD 322310 2.5HD assay- brown Sequence- based reagent RNAscope probe antiDMRT1 ACD 410481 XM_002708188.1 Sequence- based reagent RNAscope probe anti- SRY ACD 803191 AY785433.1 Sequence- based reagent RNAscope probe antiPOU5F1 ACD 513271 NM_001099957.1 Sequence- based reagent RNAscope probe antiRSPO1 ACD 488231 XM_002720657.2 Antibody Anti- DMRT1 (mouse monoclonal) Santa Cruz sc-377167 IHC (1:500) IF (1:200) WB (1:100) Antibody Anti- SOX9 (rabbit polyclonal) Francis Poulat IHC (1:500) IF (1:200) Antibody Anti- FOXL2 (rabbit polyclonal) Boulanger et al., 2014 IHC (1:500) Antibody Anti- POU5F1 (goat polyclonal) Santa Cruz sc- 8628 IHC (1:500) IF (1:200) Antibody Anti- PAX8 (rabbit polyclonal) Proteintech 10226- 1- AP IHC (1:6000) Antibody Anti- Ki67 (rabbit monoclonal) Thermo Scientific MA5- 14520 IHC (1:500) Sequence- based reagent SRY_F This paper PCR primers TGCT TACA CACC AGCC AAAC A Sequence- based reagent SRY_R This paper PCR primers TTCC TGGC CGCT CACT TTAC Sequence- based reagent DMRT1_F This paper PCR primers GGAG CCTC CCAG CACC TTA Sequence- based reagent DMRT1_R This paper PCR primers TGCA TCCT GTAC TGCG AACT CA Sequence- based reagent SOX9_F This paper PCR primers GGCT CCGA CACC GAGA ATAC AC Sequence- based reagent SOX9_R This paper PCR primers GAAC TTGT CCTC TTCG CTCT CCTT Sequence- based reagent CYP19A1_F This paper PCR primers GGAA GAAT GCAT CGAC TTGA GTT Sequence- based reagent CYP19A1_R This paper PCR primers GGGC CCAA AACC AAAT GGT Dujardin et al. eLife 2023;12:RP89284.

Techniques: Expressing, Immunostaining, Control, Quantitative RT-PCR, In Situ Hybridization, RNAscope

Journal: eLife

Article Title: DMRT1 is a testis-determining gene in rabbits and is also essential for female fertility

doi: 10.7554/elife.89284

Figure Lengend Snippet: Figure 5. Somatic markers expression and location on control and DMRT1−/− gonads at 20 dpc. Quantitative RT-PCR (RT-qPCR) analyses of (A) testicular- related differentiation genes (SOX9, AMH, DHH, and SRY) or (B) ovarian-related differentiation genes (FOXL2, CYP19A1, and RSPO1) in XY control, XY DMRT1−/−, XX DMRT1−/−, and XX control gonads (n = 4–5) at 20 dpc. Statistical analyses were performed using the non-parametric Kruskal–Wallis test, followed by a pairwise permutation test: *p-value <0.05; ns: non-significant. (C) Immunostaining of DMRT1, SOX9, and FOXL2 on XY control, XY DMRT1−/−, XX DMRT1−/−, and XX control gonad sections at 20 dpc. Scale bar = 50 µm.

Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Biological sample (Oryctolagus cuniniculus) Gonads Hypharm NZ1777 New Zealand rabbits Commercial kit RNAscope kit ACD 322310 2.5HD assay- brown Sequence- based reagent RNAscope probe antiDMRT1 ACD 410481 XM_002708188.1 Sequence- based reagent RNAscope probe anti- SRY ACD 803191 AY785433.1 Sequence- based reagent RNAscope probe antiPOU5F1 ACD 513271 NM_001099957.1 Sequence- based reagent RNAscope probe antiRSPO1 ACD 488231 XM_002720657.2 Antibody Anti- DMRT1 (mouse monoclonal) Santa Cruz sc-377167 IHC (1:500) IF (1:200) WB (1:100) Antibody Anti- SOX9 (rabbit polyclonal) Francis Poulat IHC (1:500) IF (1:200) Antibody Anti- FOXL2 (rabbit polyclonal) Boulanger et al., 2014 IHC (1:500) Antibody Anti- POU5F1 (goat polyclonal) Santa Cruz sc- 8628 IHC (1:500) IF (1:200) Antibody Anti- PAX8 (rabbit polyclonal) Proteintech 10226- 1- AP IHC (1:6000) Antibody Anti- Ki67 (rabbit monoclonal) Thermo Scientific MA5- 14520 IHC (1:500) Sequence- based reagent SRY_F This paper PCR primers TGCT TACA CACC AGCC AAAC A Sequence- based reagent SRY_R This paper PCR primers TTCC TGGC CGCT CACT TTAC Sequence- based reagent DMRT1_F This paper PCR primers GGAG CCTC CCAG CACC TTA Sequence- based reagent DMRT1_R This paper PCR primers TGCA TCCT GTAC TGCG AACT CA Sequence- based reagent SOX9_F This paper PCR primers GGCT CCGA CACC GAGA ATAC AC Sequence- based reagent SOX9_R This paper PCR primers GAAC TTGT CCTC TTCG CTCT CCTT Sequence- based reagent CYP19A1_F This paper PCR primers GGAA GAAT GCAT CGAC TTGA GTT Sequence- based reagent CYP19A1_R This paper PCR primers GGGC CCAA AACC AAAT GGT Dujardin et al. eLife 2023;12:RP89284.

Techniques: Expressing, Control, Quantitative RT-PCR, Immunostaining