mouse monoclonal anti coup tfii  (R&D Systems)

Journal: Scientific Reports

Article Title: Conditional disruption of Nr5a1 directed by Sox9-Cre impairs adrenal development

doi: 10.1038/s41598-024-63264-9

Figure Lengend Snippet: Capsule formation during embryonic development. Representative images of double-label immunofluorescence for NR5A1 (green) and NR2F2 (red) was performed on parasagittal sections of embryos from control ( a – c ) and Sox9-Cre;Nr5a1 flox/flox ( d – f ) mice at E13.5, E14.5, and E15.5. Nuclei are stained blue with DAPI. AP, adrenal primordium; K, kidney; Li, liver. Scale bars, 100 µm in a , b , d , e, 200 µm in c , f .

Article Snippet: The primary antibodies were rabbit anti-NR5A1 , rabbit anti-SOX9 (ab185230; Abcam), rabbit anti-3βHSD (KO607; Trans Genic), rabbit anti-20αHSD/AKR1C1 (OAGA00409; Aviva Systems Biology), rabbit anti-Tyrosine Hydroxylase (AB152; Millipore), rabbit anti-Cleaved Caspase-3 (9661; Cell Signaling), goat anti-GATA4 antibody (sc1237; Santa Cruz Biotechnology), mouse anti- NR2F2/COUP-TFII antibody (PP-H7147; Perseus Proteomics), and mouse anti-BrdU antibody (B8434; Sigma-Aldrich).

Techniques: Immunofluorescence, Staining

Journal: eLife

Article Title: The differentiation and integration of the hippocampal dorsoventral axis are controlled by two nuclear receptor genes

doi: 10.7554/eLife.86940

Figure Lengend Snippet: ( A ) The expression of NR2F1 ( a, d, g, j, m ) and NR2F2 ( b, e, h, k, n ) in coronal sections ( a–f ) and sagittal sections ( g–o ) of the hippocampus at postnatal month 1 (1M); representative western blots and quantitative densitometry data for the expression of NR2F1 and NR2F2 in the dorsal and ventral hippocampus at 1M ( p–q ). Data are presented as mean ± SEM. Student’s t test was used in q, * P< 0.05, ** P< 0.01; n represents separate experiments, n=3. ( B ) In coronal sections along the rostrocaudal axis ( a–f ) and sagittal sections along the lateral-medial axis ( g–l ) of the hippocampus in mutant mice, compared with that in control mice ( a–c, g–i ), the ectopic CA-like structure, indicated by the star, was observed in the ventral region in Nr2f2 gene mutant (RX Cre/+ ; Nr2f2 F/F ) mice at 1M ( d–f, j–l ). ( C ) The expression of HuB and Ctip2 in the corresponding inserted area in ( a–f ) under a high-magnification objective lens at 1M ( g–r ); compared with those of control mice ( a–c, g–i, m–o ), the duplicated HuB-positive CA3 domain, indicated by the star, and Ctip2-positive domains, indicated by the arrowhead, were specifically observed in the ventral hippocampus ( d–f, p–r ) but not in the dorsal hippocampus ( d–f, j–l ) of Nr2f2 mutant mice at 1M; Ctip2-positive AHi and MePD amygdaloid nuclei were barely observed in the Nr2f2 mutant mice, indicated by the arrows, instead of the ectopic CA domains at the prospective amygdaloid regions ( e–f, q–r ). AHi, amygdalohippocampal area; AMY, amygdala nuclei; CTX, cortex; dCA1, dorsal CA1; dCA3, dorsal CA3; dDG, dorsal dentate gyrus; dHPC, dorsal hippocampus; MePD, posterodorsal part of the medial amygdaloid nucleus; PMCo, posteromedial cortical amygdaloid nucleus; vCA1, ventral CA1; vCA3, ventral CA3; vDG, ventral dentate gyrus; vHPC, ventral hippocampus. Scale bars, ( Aa–c, Ad–f, Aj–o, Cg–r ), 100 μm; ( Ag–i, Ba–l, Ca–f ), 200 μm. Figure 1—source data 1. The Nissl staining results of the control and RX Cre/+ ; Nr2f2 F/F mutant mice at 1M (part 1); the expression of HuB and Ctip2 in the hippocampus of the control and RX Cre/+ ; Nr2f2 F/F mutant mice at 1M (part 1). Figure 1—source data 2. The expression of NR2F1 and NR2F2 in coronal sections and sagittal sections of the mouse brain at 1M (part 1); the expression of HuB and Ctip2 in the hippocampus of the control and RX Cre/+ ; Nr2f2 F/F mutant mice at 1M (part 2). Figure 1—source data 3. The expression of NR2F1 and NR2F2 in coronal sections and sagittal sections of the mouse brain at 1M (part 2). Figure 1—source data 4. The expression of NR2F1 and NR2F2 in sagittal sections of the mouse brain at 1M (part 1). Figure 1—source data 5. The expression of NR2F1 and NR2F2 in sagittal sections of the mouse brain at 1M (part 2); the Nissl staining results of the control and RX Cre/+ ; Nr2f2 F/F mutant mice at 1M (part 2). Figure 1—source data 6. The expression of NR2F1 and NR2F2 in sagittal sections of the mouse brain at 1M (part 3); western blots data for the expression of NR2F1 and NR2F2 in the dorsal and ventral hippocampus at 1M; the Nissl staining results of the control and RX Cre/+ ; Nr2f2 F/F mutant mice at 1M (part 3).

Article Snippet: The following primary antibodies were used in the study: mouse anti-NR2F1 (1:1000, R&D, Cat# PP-H8132-00), mouse anti-NR2F2 (1:2000, R&D, Cat# PP-H7147-00), rabbit anti- NR2F2 (1:2000, a gift from Dr. Zhenzhong Xu, Zhejiang University, China), rabbit anti-HuB (1:500, Abcam, Cat# ab204991), rat anti-Ctip2 (1:500, Abcam, Cat# ab18465), rabbit anti-Wfs1 (1:500, ProteinTech, Cat# 11558-1-AP), goat anti-Prox1 (1:500, R&D, Cat# AF2727), rabbit anti-Calretinin (1:500, Sigma, Cat# C7479), rabbit anti-Calbindin (1:500, Swant, Cat# CB38), mouse anti-SMI312 (1:200, Covance, Cat# SMI-312R), rabbit anti-Sox2 (1:500, Affinity BioReagents, Cat# PA1-16968), rat anti-Tbr2 (1:500, Thermo Fisher, Cat# 12-4875-82), goat anti-NeuroD1 (1:200, Santa Cruz, Cat# sc-1084), goat anti-Lhx2 (1:200, Santa Cruz, Cat# sc-19344), goat anti-Lhx5 (1:200, R&D, Cat# AF6290), goat anti-β-galactosidase (LacZ) (1:400, Biogenesis, Cat# 4600-1409), mouse anti-GFAP (1:500, Sigma, Cat# G3893), rabbit anti-Nestin (1:200, Santa Cruz, Cat# sc-20978), and goat anti-Dcx (1:500, Santa Cruz, Cat# sc-8066).

Techniques: Expressing, Western Blot, Mutagenesis, Staining

Journal: eLife

Article Title: The differentiation and integration of the hippocampal dorsoventral axis are controlled by two nuclear receptor genes

doi: 10.7554/eLife.86940

Figure Lengend Snippet: ( A ) In coronal sections along the rostrocaudal axis ( a–f ) and sagittal sections along the lateral–medial axis ( g–l ) of the hippocampus, compared with that of control mice ( a–c, g–i ), the dorsal hippocampus was shrunken, indicated by the star, in Nr2f1 gene mutant (RX Cre/+ ; Nr2f1 F/F ) mice at 1M ( d–f, j–l ). ( B ) The expression of HuB and Ctip2 in the corresponding inserted area in ( a–f ) under a high-magnification objective lens at 1M ( g–r ); compared with that of control mice ( a–c, g–i, m–o ), the HuB-positive CA3 domain was reduced in the dorsal hippocampus, especially the Ctip2-positive dorsal CA1, which was barely detected in Nr2f1 mutant mice at 1M ( d–f, j–l ), while their expression in the ventral hippocampus was comparable between the controls and mutants ( d–f, p–r ). ( C ) The expression of HuB and Wfs1 in the corresponding inserted area in ( a–f ) under a high-magnification objective lens at 1M ( g–r ); the expression of HuB and the dCA1 marker Wfs1 in the control ( a–c, g–i, m–o ) and Nr2f2 mutant mice ( d–f, j–l, p–r ) at 1M. Wfs1-positive dorsal CA1 could not be detected in Nr2f1 mutant mice at 1M, as indicated by the arrowhead. dHPC, dorsal hippocampus; HPC, hippocampus; vHPC, ventral hippocampus. Scale bars, ( Aa–l, Ba–f, Ca–f ), 200 μm; ( Bg–r, Cg–r ), 100 μm. Figure 2—source data 1. The Nissl staining results of the control and RX Cre/+ ; Nr2f1 F/F mutant mice at 1M (part 1). Figure 2—source data 2. The Nissl staining results of the control and RX Cre/+ ; Nr2f1 F/F mutant mice at 1M (part 2); the expression of HuB and Wfs1 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F mutant mice at 1M. Figure 2—source data 3. The expression of HuB and Ctip2 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F mutant mice at 1M. Figure 2—source data 4. The expression of HuB, Wfs1, and Ctip2 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F mutant mice at 1M.

Article Snippet: The following primary antibodies were used in the study: mouse anti-NR2F1 (1:1000, R&D, Cat# PP-H8132-00), mouse anti-NR2F2 (1:2000, R&D, Cat# PP-H7147-00), rabbit anti- NR2F2 (1:2000, a gift from Dr. Zhenzhong Xu, Zhejiang University, China), rabbit anti-HuB (1:500, Abcam, Cat# ab204991), rat anti-Ctip2 (1:500, Abcam, Cat# ab18465), rabbit anti-Wfs1 (1:500, ProteinTech, Cat# 11558-1-AP), goat anti-Prox1 (1:500, R&D, Cat# AF2727), rabbit anti-Calretinin (1:500, Sigma, Cat# C7479), rabbit anti-Calbindin (1:500, Swant, Cat# CB38), mouse anti-SMI312 (1:200, Covance, Cat# SMI-312R), rabbit anti-Sox2 (1:500, Affinity BioReagents, Cat# PA1-16968), rat anti-Tbr2 (1:500, Thermo Fisher, Cat# 12-4875-82), goat anti-NeuroD1 (1:200, Santa Cruz, Cat# sc-1084), goat anti-Lhx2 (1:200, Santa Cruz, Cat# sc-19344), goat anti-Lhx5 (1:200, R&D, Cat# AF6290), goat anti-β-galactosidase (LacZ) (1:400, Biogenesis, Cat# 4600-1409), mouse anti-GFAP (1:500, Sigma, Cat# G3893), rabbit anti-Nestin (1:200, Santa Cruz, Cat# sc-20978), and goat anti-Dcx (1:500, Santa Cruz, Cat# sc-8066).

Techniques: Mutagenesis, Expressing, Marker, Staining

Journal: eLife

Article Title: The differentiation and integration of the hippocampal dorsoventral axis are controlled by two nuclear receptor genes

doi: 10.7554/eLife.86940

Figure Lengend Snippet: ( A ) In coronal sections along the rostrocaudal axis, compared with control mice ( a–d ), the hippocampus was atrophic in RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice, indicated by the star, and an ectopic unknown nucleus was observed in the caudal plates, indicated by the arrowhead ( e–h ). ( B ) Compared with that of control mice ( a–c, g–i ), the expression of HuB, Ctip2, and Prox1 was decreased in the hippocampus of Nr2f1/2 double-gene mutant mice at 3 weeks postnatal (3W) ( d–f, j–l ). ( C ) Compared with that of control mice ( a–c, g–i ), the expression of HuB could not be detected in the presumptive CA3 domain, and the expression of Ctip2 or Prox1 could not be detected in the presumptive DG domain of the prospective ventral hippocampus of RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice. Scale bars, ( Aa–h ), 200 μm; ( Ba–l, Ca–l ), 100 μm; ( Bd-f (insets), Bj-l (insets)), 400 μm. Figure 3—source data 1. The Nissl staining results of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-gene mutant mice at 3W. Figure 3—source data 2. The expression of HuB and Ctip2 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-gene mutant mice at 3W. Figure 3—source data 3. The expression of HuB and Prox1 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-gene mutant mice at 3W. Figure 3—source data 4. The expression of HuB, Ctip2, and Prox1 in the hippocampus of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-gene mutant mice at 3W.

Article Snippet: The following primary antibodies were used in the study: mouse anti-NR2F1 (1:1000, R&D, Cat# PP-H8132-00), mouse anti-NR2F2 (1:2000, R&D, Cat# PP-H7147-00), rabbit anti- NR2F2 (1:2000, a gift from Dr. Zhenzhong Xu, Zhejiang University, China), rabbit anti-HuB (1:500, Abcam, Cat# ab204991), rat anti-Ctip2 (1:500, Abcam, Cat# ab18465), rabbit anti-Wfs1 (1:500, ProteinTech, Cat# 11558-1-AP), goat anti-Prox1 (1:500, R&D, Cat# AF2727), rabbit anti-Calretinin (1:500, Sigma, Cat# C7479), rabbit anti-Calbindin (1:500, Swant, Cat# CB38), mouse anti-SMI312 (1:200, Covance, Cat# SMI-312R), rabbit anti-Sox2 (1:500, Affinity BioReagents, Cat# PA1-16968), rat anti-Tbr2 (1:500, Thermo Fisher, Cat# 12-4875-82), goat anti-NeuroD1 (1:200, Santa Cruz, Cat# sc-1084), goat anti-Lhx2 (1:200, Santa Cruz, Cat# sc-19344), goat anti-Lhx5 (1:200, R&D, Cat# AF6290), goat anti-β-galactosidase (LacZ) (1:400, Biogenesis, Cat# 4600-1409), mouse anti-GFAP (1:500, Sigma, Cat# G3893), rabbit anti-Nestin (1:200, Santa Cruz, Cat# sc-20978), and goat anti-Dcx (1:500, Santa Cruz, Cat# sc-8066).

Techniques: Mutagenesis, Expressing, Staining

Journal: eLife

Article Title: The differentiation and integration of the hippocampal dorsoventral axis are controlled by two nuclear receptor genes

doi: 10.7554/eLife.86940

Figure Lengend Snippet: ( A ) The expression profiles of genes involved in hippocampal development in control and the double-mutant mice at E11.5. Data are presented as mean ± SEM. Student’s t test was used in A, * P< 0.05, ** P< 0.01; n represents separate experiments, n=3. ( B ) Compared with that of control mice ( a, b, a’, b’, a’’, b’’, i, j, i’, j’, q, r, q’, r’ ), the expression of Lhx5 was reduced in double-mutant mice at E11.5 ( c, d, c’, d’, c’’, d’’ ), E13.5 ( k, l, k’, l’ ), and E14.5 ( s, t, s’, t’ ); the expression of Lhx2 was comparable between the control and double-mutant mice at E11.5 ( e–h, e’–h’ ); and compared with that of control mice ( m, n, m’, n’, u, v, u’, v’ ), the expression of Lhx2 was increased in double-mutant mice at E13.5 ( o, p, o’, p’ ) and E14.5 ( w, x, w’, x’ ). ( C ) Compared with that of control mice ( a, b, a’, b’ ), the expression of Sox2 was normal in double-mutant mice at E14.5 ( c, d, c’, d’ ); compared with that of control mice ( e, f, e’, f’ ), the expression of Tbr2 was decreased in Nr2f mutant mice at E14.5 ( g, h, g’, h’ ); compared with that of control mice ( i, j, i’, j’ ), the expression of NeuroD1 was reduced in double-mutant mice at E14.5 ( k, l, k’, l’ ). ( D ) Quantitative analysis of Tbr2-positive cells and NeuroD1-positive cells in ( Ce’–h’ ) and ( Ci’–l’ ). Data are presented as mean ± SEM. Student’s t test was used in D, ** P< 0.01; n represents separate experiments, n=3. ( E ), In the hippocampal primordium of the early embryo, Nr2f1 is expressed dorsally in the MP, and Nr2f2 is expressed ventrally in the CH. In the mature hippocampus, the expression of Nr2f1 is higher in the dorsal hippocampus, which is related to spatial learning and memory, and the expression of Nr2f2 is mainly in the ventral hippocampus, which is associated with emotion and anxiety ( a ). Our findings support a novel molecular mechanism by which Nr2f1 and Nr2f2 may cooperate to ensure the appropriate morphogenesis and functions of the hippocampus by modulating the Lhx5-Lhx2 axis ( b ). CH, cortical hem; ChP, choroid plexus; DP, dorsal pallium; HP, hippocampal primordium; MP, medial pallium. Scale bars, ( Ba–x, Ca–l ), 200 μm; ( Ba’-x’, Ba’’-d’’, Ca’-l’ ), 100 μm. Figure 5—source data 1. The expression of Lhx5 and Lhx2 in the telencephalon of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E14.5; the expression of Tbr2 and NeuroD1 in the telencephalon of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E14.5 (part 1). Figure 5—source data 2. The expression of Sox2, Tbr2, and NeuroD1 in the telencephalon of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E14.5. Figure 5—source data 3. The expression profiles of genes involved in the hippocampal development of the control and double-mutant mice at E11.5; the expression of Lhx5 and Lhx2 in the telencephalon of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E11.5 and E13.5; the expression of Tbr2 and NeuroD1 in the telencephalon of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E14.5 (part 2); quantitative analysis of Tbr2-positive and NeuroD1-positive cells in the hippocampal primordium of the control and RX Cre/+ ; Nr2f1 F/F ; Nr2f2 F/F double-mutant mice at E14.5.

Article Snippet: The following primary antibodies were used in the study: mouse anti-NR2F1 (1:1000, R&D, Cat# PP-H8132-00), mouse anti-NR2F2 (1:2000, R&D, Cat# PP-H7147-00), rabbit anti- NR2F2 (1:2000, a gift from Dr. Zhenzhong Xu, Zhejiang University, China), rabbit anti-HuB (1:500, Abcam, Cat# ab204991), rat anti-Ctip2 (1:500, Abcam, Cat# ab18465), rabbit anti-Wfs1 (1:500, ProteinTech, Cat# 11558-1-AP), goat anti-Prox1 (1:500, R&D, Cat# AF2727), rabbit anti-Calretinin (1:500, Sigma, Cat# C7479), rabbit anti-Calbindin (1:500, Swant, Cat# CB38), mouse anti-SMI312 (1:200, Covance, Cat# SMI-312R), rabbit anti-Sox2 (1:500, Affinity BioReagents, Cat# PA1-16968), rat anti-Tbr2 (1:500, Thermo Fisher, Cat# 12-4875-82), goat anti-NeuroD1 (1:200, Santa Cruz, Cat# sc-1084), goat anti-Lhx2 (1:200, Santa Cruz, Cat# sc-19344), goat anti-Lhx5 (1:200, R&D, Cat# AF6290), goat anti-β-galactosidase (LacZ) (1:400, Biogenesis, Cat# 4600-1409), mouse anti-GFAP (1:500, Sigma, Cat# G3893), rabbit anti-Nestin (1:200, Santa Cruz, Cat# sc-20978), and goat anti-Dcx (1:500, Santa Cruz, Cat# sc-8066).

Techniques: Expressing, Mutagenesis