Journal: Cerebral Cortex (New York, NY)

Article Title: Dlx1/2 are Central and Essential Components in the Transcriptional Code for Generating Olfactory Bulb Interneurons

doi: 10.1093/cercor/bhz018

Figure Lengend Snippet: Neural progenitors accumulate in the dLGE of Dlx1/2−/− mice at E16.5. (A–B′) Compared with controls, there were more GSX2+ and ASCL1+ progenitors in the d/vLGE of Dlx1/2−/− mice (arrows). (C, C′) ISL1 expression was increased in the mutant dLGE (arrows). (E, E′) SP8 was lost in the mutant dLGE (arrows). (F–G′) EBF1 was not expressed in the control and mutant dLGE, but the pan EBF antibody immunostaining showed that EBF3 was expressed in the mutant dLGE (arrows). (I–K′) SP9 expression was reduced in the dLGE; most SP9+ cells coexpressed the immature MSN marker BCL11b (arrows in the inset of K′). (D, H, L) Quantification of cell numbers in the dLGE. Scale bars: 200 μm in K′ for A–K′; 20 μm in the inset of K′.

Article Snippet: The following primary antibodies were used: goat anti-SP8 (1:3 000, Santa Cruz, sc-104 661), rabbit anti-SP9 (1:500) ( Zhang et al. 2016 ), chicken anti-GFP (1:3 000, Aves Labs, GFP-1020), goat anti-DCX (1:1 000, Santa Cruz, sc-8066), rabbit anti-DCX (1:1 000, Abcam, ab18723), mouse anti-NeuN (1:500, Millipore, MAB-377), rabbit anti-ASCL1 (1:2 000, Cosmo Bio, SK-T01-003), rabbit anti-GSX2 (1:2 000, Millipore, ABN162), rabbit anti-OLIG2 (1:500, Millipore, AB9610), rabbit anti-Ki67 (1:500, Vector Labs, VP-K451), rabbit anti-PAX6 (1:2 000, MBL, PD022), goat anti-EBF (1:200, Santa Cruz, sc-15 888), rabbit anti-EBF1 (1:5 000, Millipore, AB10523), goat anti-SOX2 (1:500, Santa Cruz, sc-17 320), rabbit anti-ISL1 (1:500, Abcam, ab20670), rabbit anti-GFAP (1:500, Dako, Z0334), rabbit anti-EOMES (TBR2, 1:500, Abcam, ab23345), rat anti-BCL11b (1:1 000, Abcam, ab18465), and guinea pig anti DLX2 (1:2000) ( Kuwajima et al. 2006 ; Hansen et al. 2013 ).

Techniques: Expressing, Mutagenesis, Immunostaining, Marker

Journal: PLoS Pathogens

Article Title: EBNA2 Drives Formation of New Chromosome Binding Sites and Target Genes for B-Cell Master Regulatory Transcription Factors RBP-jκ and EBF1

doi: 10.1371/journal.ppat.1005339

Figure Lengend Snippet: EREB2.5 cells were treated with (+) or without (-) estradiol (E2) for 24 or 48 hrs and then assayed by ChIP for binding to EBF1 ( A and B ), RBP-jκ ( C and D ), or EBNA2 ( E and F ) at cellular ( A, C, E ) or EBV genome sites ( B, D, F ). Actin genomic region (cellular) or Qp (EBV) was used as negative binding control for EBF1, RBP-jκ, ορ EBNA2 ChIP. (G) ChIP binding for CTCF, PU.1, or PAX5 in EREB2.5 cells treated (blue) or untreated (red) with E2 for 48 hrs. PPP1R1B or KCTD17 genomic region was negative binding control PU.1 or PAX5, respectively. Asterisk indicates p < 0.05. (H) Western blot showing protein levels for EBF1, RBP-jκ, EBNA2, Actin, and CTCF in EREB2.5 cells at 24 and 48 hrs after E2 withdrawal.

Article Snippet: Rabbit polyclonal anti-EBF1 (EMD Millipore AB10523), RBP- jκ (Abcam AB25949), CTCF (EMD Millipore 07–729), PARP-1 (Alexis 210-302-R100), GAPDH (Cell signaling 2118); Goat polyclonal anti-EBF1 (R&D System AF5165); Sheep polyclonal anti-EBNA3C (Exalpha F125P); Rat monoclonal anti-EBNA2 (EMD Millipore MABE8); Mouse monoclonal anti-LMP1 (DAKO M0897); Actin-Peroxidase antibody (Sigma A3854).

Techniques: Binding Assay, Western Blot

Journal: Clinical and Translational Medicine

Article Title: Long noncoding RNA IL6‐AS1 is highly expressed in chronic obstructive pulmonary disease and is associated with interleukin 6 by targeting miR‐149‐5p and early B‐cell factor 1

doi: 10.1002/ctm2.479

Figure Lengend Snippet: IL6‐AS1 interacts with EBF1 to regulate IL‐6 expression by affecting the binding of EBF1 to the IL‐6 promoter. (A) RNA pull‐down assay using IL6‐AS1 sense and antisense RNAs in HBF cells, followed by silver staining. Black arrow indicates EBF1. The interaction between IL6‐AS1 and EBF1 was confirmed by Western blotting with the extract of the RNA pull‐down assay. ILF2 and GPR94 were also detected. (B) RIP‐qPCR analysis using an anti‐EBF1 antibody showed that IL6‐AS1 interacts with endogenous EBF1 in HBF and HFL1 cells. U1 was used as the negative control (two‐way ANOVA, n = 3 biological replicates). (C) RIP‐qPCR analysis with an anti‐EBF1 antibody was conducted after overexpression of IL6‐AS1 in HBF and HFL1 cells (two‐way ANOVA, n = 3 biological replicates). (D) Schematic representation of potential EBF1 binding sites on the IL‐6 promoter. As promoters considered to be located 2000 bp upstream of the transcription start site (TSS), we designed six pairs of chromatin immunoprecipitation (ChIP)‐qPCR primers to cover the IL‐6 promoter region, with the fifth and sixth primers containing two EBF1 binding sites. (E) ChIP‐PCR analysis of EBF1 occupancy on the IL‐6 promoter in HBF and HLF cells. IgG was used as the negative control (two‐way ANOVA, n = 3 biological replicates). (F) ChIP‐PCR analysis of EBF1 occupancy on the IL‐6 promoter after transfection with an IL6‐AS1 silencer (SSIL6‐AS1) in HBF and HLF cells (two‐way ANOVA, n = 3 biological replicates). (G) Luciferase activity in the IL‐6 promoter following cotransfection of the IL6‐AS1 silencer (SSIL6‐AS1) or IL6‐AS1 overexpression vector (two‐way ANOVA, n = 4 biological replicates). (H) Luciferase activity in the IL‐6 promoter following cotransfection of an EBF1 small interfering RNA (siRNA) (siEBF1‐1) and EBF1 overexpression vector (two‐way ANOVA, n = 4 biological replicates). (I) Schematic representation of the two mutation sequences of potential EBF1 binding sites on the IL‐6 promoter. (J) Luciferase activity in the IL‐6 promoter following transfection with a reporter containing wild‐type or mutant IL‐6 promoter (one‐way ANOVA, n = 4 biological replicates). (K and L) qRT‐PCR and ELISA analysis of IL‐6 expression after transfection with two EBF1 siRNAs in HBF cells (K) or HLF cells (L) (one‐way ANOVA, n = 4 biological replicates). (M and N) qRT‐PCR and ELISA analysis of IL‐6 expression after overexpression of EBF1 in HBF cells (M) or HLF cells (N) (one‐way ANOVA, n = 4 biological replicates). (O and P) Expression of IL‐6 in HBF cells following cotransfection with IL6‐AS1 overexpression vector and EBF1 siRNA (SiEBF1‐1), determined by qRT‐PCR (O) and ELISA (P) (one‐way ANOVA, n = 5 biological replicates). (Q and R) Expression of IL‐6 in HBF cells following cotransfection with an IL6‐AS1 Smart Silencer (SSIL6‐AS1) and EBF1 overexpression vector, determined by qRT‐PCR (Q) and ELISA (R) (one‐way ANOVA, n = 5 biological replicates). Error bars represent means ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The cells were then incubated with rabbit anti‐human monoclonal antibody against EBF1 (1:200, ab108369, Abcam) overnight at 4°C, followed by incubation with Alexa Fluor 488 goat anti‐rabbit IgG (H+L) (1:500, Molecular Probes, Invitrogen) for 60 min.

Techniques: Expressing, Binding Assay, Pull Down Assay, Silver Staining, Western Blot, Negative Control, Over Expression, Chromatin Immunoprecipitation, Transfection, Luciferase, Activity Assay, Cotransfection, Plasmid Preparation, Small Interfering RNA, Mutagenesis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Translational Medicine

Article Title: Long noncoding RNA IL6‐AS1 is highly expressed in chronic obstructive pulmonary disease and is associated with interleukin 6 by targeting miR‐149‐5p and early B‐cell factor 1

doi: 10.1002/ctm2.479

Figure Lengend Snippet: Two hairpin structures of IL6‐AS1 can bind to the EBF1 protein. (A) Two computational methods (centroid plain structure and minimum free energy plain structure) were used to compute secondary structures of IL6‐AS1 using the RNAfold database ( http://rna.tbi.univie.ac.at/cgi‐bin/RNAWebSuite/RNAfold.cgi ). The sequence regions in red are the stable regions in the IL6‐AS1 secondary structure and the blue sequence regions are the unstable regions. The hairpin structures in the red and blue boxes are the potential binding sites of the EBF1 transcription factor. The structures in the black boxes are the highly conserved stem‐loops. (B) RIP‐qPCR analysis with an anti‐EBF1 antibody after transfection with wild‐type or truncated IL6‐AS1 (49‐332aa, 333‐568aa, 694‐1266aa) in HBF cells (two‐way ANOVA, n = 3 biological replicates). (C and D) Schematic representation of the EBF1 binding motif (C) and the two mutation sequences of potential EBF1 binding sites (D) in IL6‐AS1. (E and F) RIP‐qPCR analysis with an anti‐EBF1 antibody after transfection with wild‐type or mutant IL6‐AS1 in HBF cells (E) and HFL1 cells (F) (two‐way ANOVA, n = 4 biological replicates). (G and H) qRT‐PCR (G) and ELISA (H) analysis of IL‐6 expression after transfection with wild‐type or mutant IL6‐AS1 in HBF cells and HFL1 cells (one‐way ANOVA, n = 3 biological replicates). (I and J) HBF cells were cotransfected with wild‐type or mutant IL6‐AS1 and SSIL6‐AS1 and IL‐6 expression was determined by qRT‐PCR (I) and ELISA (J) (one‐way ANOVA, n = 4 biological replicates). (K and L) HBF cells were cotransfected with wild‐type or mutant IL6‐AS1 and siEBF1‐1 and IL‐6 expression was determined by qRT‐PCR (K) and ELISA (L) (one‐way ANOVA, n = 4 biological replicates). Error bars represent means ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The cells were then incubated with rabbit anti‐human monoclonal antibody against EBF1 (1:200, ab108369, Abcam) overnight at 4°C, followed by incubation with Alexa Fluor 488 goat anti‐rabbit IgG (H+L) (1:500, Molecular Probes, Invitrogen) for 60 min.

Techniques: Sequencing, Binding Assay, Transfection, Mutagenesis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Clinical and Translational Medicine

Article Title: Long noncoding RNA IL6‐AS1 is highly expressed in chronic obstructive pulmonary disease and is associated with interleukin 6 by targeting miR‐149‐5p and early B‐cell factor 1

doi: 10.1002/ctm2.479

Figure Lengend Snippet: IL6‐AS1 promotes the modification of the histones H3K4me3 and H3K27ac on the interleukin (IL) 6 promoter, probably through EBF1. (A) Schematic representation of the putative modification markers, H3K27ac, H3K4me1, and H3K4me3 upstream of IL‐6 from the ENCODE database ( https://genome.ucsc.edu/ENCODE/ ). (B–D) ChIP‐PCR analysis of H3K27ac (B), H3K4me1 (C), and H3K4me3 (D) on the IL‐6 promoter in HBF cells (two‐way ANOVA, n = 3 biological replicates). (E–H) HBF cells were transfected with SSIL6‐AS1 and assessed for H3K27ac (E), H3K4me3 (F), H3K4me1 (G) and RNA polymerase II (H) on the IL‐6 promoter by ChIP‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). (I–L) HBF cells were transfected with siEBF1‐1 and assessed for H3K27ac (I), H3K4me1 (J), H3K4me3 (K), and RNA polymerase II (L) on the IL‐6 promoter by chromatin immunoprecipitation‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). (M and N) HBF cells were cotransfected with siEBF1‐1 and IL6‐AS1 overexpression vector and assessed for H3K27ac (M) and H3K4me3 (N) on the IL‐6 promoter by ChIP‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). Error bars represent means ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The cells were then incubated with rabbit anti‐human monoclonal antibody against EBF1 (1:200, ab108369, Abcam) overnight at 4°C, followed by incubation with Alexa Fluor 488 goat anti‐rabbit IgG (H+L) (1:500, Molecular Probes, Invitrogen) for 60 min.

Techniques: Modification, Transfection, Chromatin Immunoprecipitation, Over Expression, Plasmid Preparation

Journal: Clinical and Translational Medicine

Article Title: Long noncoding RNA IL6‐AS1 is highly expressed in chronic obstructive pulmonary disease and is associated with interleukin 6 by targeting miR‐149‐5p and early B‐cell factor 1

doi: 10.1002/ctm2.479

Figure Lengend Snippet: Two mechanisms that synergize to promote interleukin (IL) 6 expression. (A and B) qRT‐PCR analysis of IL‐6 expression after cotransfection of HBF cells (A) and HFL1 cells (B) with wild‐type IL6‐AS1 or EBF1‐mutant IL6‐AS1 and miRNA‐mutant IL6‐AS1 (one‐way ANOVA, n = 5 biological replicates). (C and D) ELISA analysis of IL‐6 secretion after cotransfection of HBF cells (C) and HFL1 cells (D) with wild‐type IL6‐AS1 or EBF1‐mutant IL6‐AS1 and miRNA‐mutant IL6‐AS1 (one‐way ANOVA, n = 5 biological replicates). (E) HBF and HFL1 cells were exposed to LPS (500 ng/ml), cigarette smoke extract (CSE, 0.015%), PM 2.5 (2 μg/ml), IL17A (200 ng/ml), or nicotine (10 μM) for 24 h. qRT‐PCR analysis of IL6‐AS1 expression (two‐way ANOVA, n = 3 biological replicates). (F and G) qRT‐PCR and ELISA analysis of IL‐6 expression in wild‐type IL6‐AS1 cells, EBF1‐mutant IL6‐AS1 or miRNA‐mutant IL6‐AS1 cells after exposure to lipopolysaccharide (500 ng/ml) for 24 h in HFL1 (F) and HBF cells (G) (two‐way ANOVA, n = 3 biological replicates). (H) Correlation analysis of gene expression between IL6‐AS1/IL‐6, IL6‐AS1/miR‐149‐5p, and IL6/miR‐149‐5p from RNA‐seq results. (I) Correlation analysis between the expression of IL6‐AS1 and FEV1% in verified samples; correlation analysis between the expression of IL6‐AS1 and GOLD stage in verified samples. (J) Correlation analysis of gene expression between IL6‐AS1/IL‐6 in GSE38974 and GSE76925. (K) Correlation analysis between the expression of IL6‐AS1 and GOLD stage in GSE38974 and GSE76925. (L) Overview of the involvement of IL6‐AS1 in chronic obstructive pulmonary disease (COPD). Schematic representation of the mechanisms by which IL6‐AS1 regulates IL‐6 expression: promoting transcription and affecting histone modification by direct binding with EBF1 in the nucleus, and stabilizing IL‐6 mRNA by acting as a competing endogenous RNA (ceRNA) for miR‐149‐5p in the cytoplasm. Error bars represent the mean ± SD. * p < 0.05 and ** p < 0.01

Article Snippet: The cells were then incubated with rabbit anti‐human monoclonal antibody against EBF1 (1:200, ab108369, Abcam) overnight at 4°C, followed by incubation with Alexa Fluor 488 goat anti‐rabbit IgG (H+L) (1:500, Molecular Probes, Invitrogen) for 60 min.

Techniques: Expressing, Quantitative RT-PCR, Cotransfection, Mutagenesis, Enzyme-linked Immunosorbent Assay, RNA Sequencing Assay, Modification, Binding Assay

Journal: Cerebral Cortex (New York, NY)

Article Title: Dlx1/2 are Central and Essential Components in the Transcriptional Code for Generating Olfactory Bulb Interneurons

doi: 10.1093/cercor/bhz018

Figure Lengend Snippet: SP8, SP9, and DLX2 are coexpressed in the E16.5 dLGE SVZ. (A, B) GSX2 was strongly expressed in the dLGE VZ and SVZ1; only a few GSX2+ cells were in the SVZ2, whereas DLX2 was mainly expressed in the SVZ1&2. Note weak expression of GSX2 in the vLGE. (C) The section was stained with DAPI. (D, E) Most ASCL1+ cells expressed DLX2. (F–I) DLX2 was strongly expressed in the SVZ1 and relatively weakly expressed in the SVZ2 and DLX2 expression began before SP8/9 expression. Most cells in the SVZ2 expressed DLX2/SP8/SP9. (J, K) Quantification of DLX2+ cells that expressed SP8, SP9, or SP8/9 in the dLGE SVZ1 and SVZ2 at E16.5. Scale bar: 100 μm in G for (A–I).

Article Snippet: The following primary antibodies were used: goat anti-SP8 (1:3 000, Santa Cruz, sc-104 661), rabbit anti-SP9 (1:500) ( Zhang et al. 2016 ), chicken anti-GFP (1:3 000, Aves Labs, GFP-1020), goat anti-DCX (1:1 000, Santa Cruz, sc-8066), rabbit anti-DCX (1:1 000, Abcam, ab18723), mouse anti-NeuN (1:500, Millipore, MAB-377), rabbit anti-ASCL1 (1:2 000, Cosmo Bio, SK-T01-003), rabbit anti-GSX2 (1:2 000, Millipore, ABN162), rabbit anti-OLIG2 (1:500, Millipore, AB9610), rabbit anti-Ki67 (1:500, Vector Labs, VP-K451), rabbit anti-PAX6 (1:2 000, MBL, PD022), goat anti-EBF (1:200, Santa Cruz, sc-15 888), rabbit anti-EBF1 (1:5 000, Millipore, AB10523), goat anti-SOX2 (1:500, Santa Cruz, sc-17 320), rabbit anti-ISL1 (1:500, Abcam, ab20670), rabbit anti-GFAP (1:500, Dako, Z0334), rabbit anti-EOMES (TBR2, 1:500, Abcam, ab23345), rat anti-BCL11b (1:1 000, Abcam, ab18465), and guinea pig anti DLX2 (1:2000) ( Kuwajima et al. 2006 ; Hansen et al. 2013 ).

Techniques: Expressing, Staining

Journal: Cerebral Cortex (New York, NY)

Article Title: Dlx1/2 are Central and Essential Components in the Transcriptional Code for Generating Olfactory Bulb Interneurons

doi: 10.1093/cercor/bhz018

Figure Lengend Snippet: Neural progenitors accumulate in the dLGE of Dlx1/2−/− mice at E16.5. (A–B′) Compared with controls, there were more GSX2+ and ASCL1+ progenitors in the d/vLGE of Dlx1/2−/− mice (arrows). (C, C′) ISL1 expression was increased in the mutant dLGE (arrows). (E, E′) SP8 was lost in the mutant dLGE (arrows). (F–G′) EBF1 was not expressed in the control and mutant dLGE, but the pan EBF antibody immunostaining showed that EBF3 was expressed in the mutant dLGE (arrows). (I–K′) SP9 expression was reduced in the dLGE; most SP9+ cells coexpressed the immature MSN marker BCL11b (arrows in the inset of K′). (D, H, L) Quantification of cell numbers in the dLGE. Scale bars: 200 μm in K′ for A–K′; 20 μm in the inset of K′.

Article Snippet: The following primary antibodies were used: goat anti-SP8 (1:3 000, Santa Cruz, sc-104 661), rabbit anti-SP9 (1:500) ( Zhang et al. 2016 ), chicken anti-GFP (1:3 000, Aves Labs, GFP-1020), goat anti-DCX (1:1 000, Santa Cruz, sc-8066), rabbit anti-DCX (1:1 000, Abcam, ab18723), mouse anti-NeuN (1:500, Millipore, MAB-377), rabbit anti-ASCL1 (1:2 000, Cosmo Bio, SK-T01-003), rabbit anti-GSX2 (1:2 000, Millipore, ABN162), rabbit anti-OLIG2 (1:500, Millipore, AB9610), rabbit anti-Ki67 (1:500, Vector Labs, VP-K451), rabbit anti-PAX6 (1:2 000, MBL, PD022), goat anti-EBF (1:200, Santa Cruz, sc-15 888), rabbit anti-EBF1 (1:5 000, Millipore, AB10523), goat anti-SOX2 (1:500, Santa Cruz, sc-17 320), rabbit anti-ISL1 (1:500, Abcam, ab20670), rabbit anti-GFAP (1:500, Dako, Z0334), rabbit anti-EOMES (TBR2, 1:500, Abcam, ab23345), rat anti-BCL11b (1:1 000, Abcam, ab18465), and guinea pig anti DLX2 (1:2000) ( Kuwajima et al. 2006 ; Hansen et al. 2013 ).

Techniques: Expressing, Mutagenesis, Immunostaining, Marker

Journal: Cerebral Cortex (New York, NY)

Article Title: Dlx1/2 are Central and Essential Components in the Transcriptional Code for Generating Olfactory Bulb Interneurons

doi: 10.1093/cercor/bhz018

Figure Lengend Snippet: More subpallial neural progenitors are in the cortex of Pax6Sey/Sey; Dlx1/2−/− Mice at E16.5. (A–B″) Immunostaining of PAX6 and DLX2 confirmed the genotypes of mice used. (C–C″) EOMES expression was greatly reduced in both Pax6Sey/Seyand Pax6Sey/Sey; Dlx1/2−/− mice. (D–F″) GSX2+, OLIG2+, and ASCL1+ progenitors ectopically accumulated in the neocortex of Pax6Sey/Sey mice; their expression increased further in Pax6Sey/Sey; Dlx1/2−/− mice. (G–I″) The ectopic SP8/9 cortical expression in the Pax6Sey/Sey mice was reduced in the Pax6Sey/Sey; Dlx1/2−/− cortex; furthermore, the SP8/9+ cells showed a scattered pattern. (J–L″) Most SP9/8+ cells coexpressed BCL11b in the cortex of Pax6Sey/Sey; Dlx1/2−/− mice. (M–N″) ISL1 and EBF1 expression was not observed in the neocortex of these mutants. Scale bars: 70 μm in L″ for L–L″; 250 μm in M″ for A–F″, H–K″, M–M″; 500 μm in N″ for G–G″, N–N″.

Article Snippet: The following primary antibodies were used: goat anti-SP8 (1:3 000, Santa Cruz, sc-104 661), rabbit anti-SP9 (1:500) ( Zhang et al. 2016 ), chicken anti-GFP (1:3 000, Aves Labs, GFP-1020), goat anti-DCX (1:1 000, Santa Cruz, sc-8066), rabbit anti-DCX (1:1 000, Abcam, ab18723), mouse anti-NeuN (1:500, Millipore, MAB-377), rabbit anti-ASCL1 (1:2 000, Cosmo Bio, SK-T01-003), rabbit anti-GSX2 (1:2 000, Millipore, ABN162), rabbit anti-OLIG2 (1:500, Millipore, AB9610), rabbit anti-Ki67 (1:500, Vector Labs, VP-K451), rabbit anti-PAX6 (1:2 000, MBL, PD022), goat anti-EBF (1:200, Santa Cruz, sc-15 888), rabbit anti-EBF1 (1:5 000, Millipore, AB10523), goat anti-SOX2 (1:500, Santa Cruz, sc-17 320), rabbit anti-ISL1 (1:500, Abcam, ab20670), rabbit anti-GFAP (1:500, Dako, Z0334), rabbit anti-EOMES (TBR2, 1:500, Abcam, ab23345), rat anti-BCL11b (1:1 000, Abcam, ab18465), and guinea pig anti DLX2 (1:2000) ( Kuwajima et al. 2006 ; Hansen et al. 2013 ).

Techniques: Immunostaining, Expressing

Journal: Cerebral Cortex (New York, NY)

Article Title: Dlx1/2 are Central and Essential Components in the Transcriptional Code for Generating Olfactory Bulb Interneurons

doi: 10.1093/cercor/bhz018

Figure Lengend Snippet: Neuronal differentiation of stem/progenitors in the adult SVZ (P50) of hGFAP-Cre; Dlx1/2F/− conditional knockout (Dlx1/2-CKO) mice was blocked. (A–L′) DLX2+, SP8+, SP9+, DLX2+/DCX+, SP8+/DCX+, and SP9+/DCX+ cells were greatly reduced whereas GSX2+, Ki67+, ASCL1+, GSX2+/DCX+, Ki67+/DCX+, and ASCL1+/DCX+ cells were significantly increased in the SVZ of Dlx1/2-CKO mice compared with wild type control mice at P50. (M–R) Quantification of these experiments. Scale bar: 50 μm in L′ for A–L′.

Article Snippet: The following primary antibodies were used: goat anti-SP8 (1:3 000, Santa Cruz, sc-104 661), rabbit anti-SP9 (1:500) ( Zhang et al. 2016 ), chicken anti-GFP (1:3 000, Aves Labs, GFP-1020), goat anti-DCX (1:1 000, Santa Cruz, sc-8066), rabbit anti-DCX (1:1 000, Abcam, ab18723), mouse anti-NeuN (1:500, Millipore, MAB-377), rabbit anti-ASCL1 (1:2 000, Cosmo Bio, SK-T01-003), rabbit anti-GSX2 (1:2 000, Millipore, ABN162), rabbit anti-OLIG2 (1:500, Millipore, AB9610), rabbit anti-Ki67 (1:500, Vector Labs, VP-K451), rabbit anti-PAX6 (1:2 000, MBL, PD022), goat anti-EBF (1:200, Santa Cruz, sc-15 888), rabbit anti-EBF1 (1:5 000, Millipore, AB10523), goat anti-SOX2 (1:500, Santa Cruz, sc-17 320), rabbit anti-ISL1 (1:500, Abcam, ab20670), rabbit anti-GFAP (1:500, Dako, Z0334), rabbit anti-EOMES (TBR2, 1:500, Abcam, ab23345), rat anti-BCL11b (1:1 000, Abcam, ab18465), and guinea pig anti DLX2 (1:2000) ( Kuwajima et al. 2006 ; Hansen et al. 2013 ).

Techniques: Knock-Out