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Proteintech dach1 protein

Fig. 2 <t>DACH1</t> deletion PCa enhances AR signaling. A Interrogation of human PCa gene expression data [26], showing candidate genetic drivers ERG, ETV1/ETV4/FLI1, SPOP, FOXA1, and unknown. Samples with DACH1 homozygous (deep) genetic deletions (29/333) are shown as an additional subtype. The AR score (the average of the AR target gene expression) refers to a group of AR-responsive genes [26], and together with the expression Z-score of the AR target genes, are shown as colorimetric scales. The AR score-based gene names are shown. The androgen receptor (AR) activity, inferred by the induction of AR target genes, was increased in DACH1 homozygous (‘deep’) deletion PCa compared with normal (P = 2 × 10−5 by t-test) and ERG mutation groups (P = 0.003 by t-test). B AR mRNA and AR protein levels, shown for each DACH1 deletion sample, were not signiﬁcantly different. C The iCluster [29], mRNA cluster, and SCNA (somatic copy-number alteration), and DNA methylation status are shown for the PCa classiﬁed by the corresponding gene deletion subtypes. D DACH1 homozygous deletions were enriched for iCluster 2 and 3 [29], mRNA cluster 2 (P = 0.0003 by Fisher exact test, SCNA (“more” somatic copy-number alteration, P = 0.0004 by Fisher exact test), but not for DNA methylation.

Dach1 Protein, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses."

Article Title: The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses.

Journal: Oncogene

doi: 10.1038/s41388-023-02668-9

Fig. 2 DACH1 deletion PCa enhances AR signaling. A Interrogation of human PCa gene expression data [26], showing candidate genetic drivers ERG, ETV1/ETV4/FLI1, SPOP, FOXA1, and unknown. Samples with DACH1 homozygous (deep) genetic deletions (29/333) are shown as an additional subtype. The AR score (the average of the AR target gene expression) refers to a group of AR-responsive genes [26], and together with the expression Z-score of the AR target genes, are shown as colorimetric scales. The AR score-based gene names are shown. The androgen receptor (AR) activity, inferred by the induction of AR target genes, was increased in DACH1 homozygous (‘deep’) deletion PCa compared with normal (P = 2 × 10−5 by t-test) and ERG mutation groups (P = 0.003 by t-test). B AR mRNA and AR protein levels, shown for each DACH1 deletion sample, were not signiﬁcantly different. C The iCluster [29], mRNA cluster, and SCNA (somatic copy-number alteration), and DNA methylation status are shown for the PCa classiﬁed by the corresponding gene deletion subtypes. D DACH1 homozygous deletions were enriched for iCluster 2 and 3 [29], mRNA cluster 2 (P = 0.0003 by Fisher exact test, SCNA (“more” somatic copy-number alteration, P = 0.0004 by Fisher exact test), but not for DNA methylation.

Figure Legend Snippet: Fig. 2 DACH1 deletion PCa enhances AR signaling. A Interrogation of human PCa gene expression data [26], showing candidate genetic drivers ERG, ETV1/ETV4/FLI1, SPOP, FOXA1, and unknown. Samples with DACH1 homozygous (deep) genetic deletions (29/333) are shown as an additional subtype. The AR score (the average of the AR target gene expression) refers to a group of AR-responsive genes [26], and together with the expression Z-score of the AR target genes, are shown as colorimetric scales. The AR score-based gene names are shown. The androgen receptor (AR) activity, inferred by the induction of AR target genes, was increased in DACH1 homozygous (‘deep’) deletion PCa compared with normal (P = 2 × 10−5 by t-test) and ERG mutation groups (P = 0.003 by t-test). B AR mRNA and AR protein levels, shown for each DACH1 deletion sample, were not signiﬁcantly different. C The iCluster [29], mRNA cluster, and SCNA (somatic copy-number alteration), and DNA methylation status are shown for the PCa classiﬁed by the corresponding gene deletion subtypes. D DACH1 homozygous deletions were enriched for iCluster 2 and 3 [29], mRNA cluster 2 (P = 0.0003 by Fisher exact test, SCNA (“more” somatic copy-number alteration, P = 0.0004 by Fisher exact test), but not for DNA methylation.

Techniques Used: Gene Expression, Targeted Gene Expression, Expressing, Activity Assay, Mutagenesis, DNA Methylation Assay

Fig. 3 Prostate-speciﬁc Dach1 gene deletion promotes prostate hyperplasia and dysplasia in OncoMice (15 weeks). A Schematic representation of transgenes integrated into mice. B Representative immunohistochemistry for Dach1, with data quantitated as mean ± standard error of the mean (SEM) for N = 20 (4 separate mice, with 5 views per mouse, in each group). C Blinded quantitative histology grading of prostate of multigenic mice at 15 weeks. Data are shown as mean ± SEM for N = 15 (5 separate mice, with 3 prostate areas [anterior, ventral, lateral] per mouse) in each group). H&E staining demonstrates the presence of a focal atypical intraductal proliferation in Dach1−/−prostate, compatible with prostatic intraepithelial neoplasia (PIN). Representative immunohistochemistry with results shown as mean ± SEM for Ki-67 (n = 20, 4 separate mice for each genotype, 5 views per mouse) (D), Beclin 1 (n = 9, 3 separate mice for each genotype, 3 views per mouse) (E); and AR (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 12 for Dach1ﬂ/ﬂmice, 3 separate mice, 2 views for one mouse and 5 views for other two mice) (F). Scale bars, 50 μm. A Student’s t test was performed for all comparisons.

Figure Legend Snippet: Fig. 3 Prostate-speciﬁc Dach1 gene deletion promotes prostate hyperplasia and dysplasia in OncoMice (15 weeks). A Schematic representation of transgenes integrated into mice. B Representative immunohistochemistry for Dach1, with data quantitated as mean ± standard error of the mean (SEM) for N = 20 (4 separate mice, with 5 views per mouse, in each group). C Blinded quantitative histology grading of prostate of multigenic mice at 15 weeks. Data are shown as mean ± SEM for N = 15 (5 separate mice, with 3 prostate areas [anterior, ventral, lateral] per mouse) in each group). H&E staining demonstrates the presence of a focal atypical intraductal proliferation in Dach1−/−prostate, compatible with prostatic intraepithelial neoplasia (PIN). Representative immunohistochemistry with results shown as mean ± SEM for Ki-67 (n = 20, 4 separate mice for each genotype, 5 views per mouse) (D), Beclin 1 (n = 9, 3 separate mice for each genotype, 3 views per mouse) (E); and AR (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 12 for Dach1ﬂ/ﬂmice, 3 separate mice, 2 views for one mouse and 5 views for other two mice) (F). Scale bars, 50 μm. A Student’s t test was performed for all comparisons.

Techniques Used: Immunohistochemistry, Staining

Fig. 4 Prostate-speciﬁc Dach1 gene deletion in TRAMP mice induces PIN lesions with increased TGFβ activity. Genome-wide expression analysis of TRAMP Dach1+/+ vs. Dach1−/−PIN lesions was analyzed for enrichment of known targets of upstream regulators using Ingenuity Pathway Analysis (IPA) and represented as (A) barplot was calculated by IPA activation Z-score labeled and as (B) bubble plot with size of the bubbles proportional to –log10 p values. C IHC was conducted for SMAD activation using SMAD2P, quantitated and shown as (D) mean ± SEM (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 10 for Dach1ﬂ/ﬂmice, 2 separate mice, 5 views per mouse). E–G Western blot of either PCa cell lines for the presence of DACH1 (E, F) or (G) TGFβ-treated (10 ng/ml for 24 h) PC3 cells illustrating induction of nuclear vimentin and cytoplasmic cyclin D1. Protein loading controls are β-tubulin (a marker of cytoplasmic proteins) and Lamin B1 (a marker for nuclear protein enrichment). H Microarray-based gene expression analysis of PC3 cells stably expressing DACH1, showing restraint of genes mediating TGFβ signaling (shown with blue arrows), including reduction of TGFB2 and TGFBR2 [33].

Figure Legend Snippet: Fig. 4 Prostate-speciﬁc Dach1 gene deletion in TRAMP mice induces PIN lesions with increased TGFβ activity. Genome-wide expression analysis of TRAMP Dach1+/+ vs. Dach1−/−PIN lesions was analyzed for enrichment of known targets of upstream regulators using Ingenuity Pathway Analysis (IPA) and represented as (A) barplot was calculated by IPA activation Z-score labeled and as (B) bubble plot with size of the bubbles proportional to –log10 p values. C IHC was conducted for SMAD activation using SMAD2P, quantitated and shown as (D) mean ± SEM (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 10 for Dach1ﬂ/ﬂmice, 2 separate mice, 5 views per mouse). E–G Western blot of either PCa cell lines for the presence of DACH1 (E, F) or (G) TGFβ-treated (10 ng/ml for 24 h) PC3 cells illustrating induction of nuclear vimentin and cytoplasmic cyclin D1. Protein loading controls are β-tubulin (a marker of cytoplasmic proteins) and Lamin B1 (a marker for nuclear protein enrichment). H Microarray-based gene expression analysis of PC3 cells stably expressing DACH1, showing restraint of genes mediating TGFβ signaling (shown with blue arrows), including reduction of TGFB2 and TGFBR2 [33].

Techniques Used: Activity Assay, Genome Wide, Expressing, Activation Assay, Labeling, Western Blot, Marker, Protein Enrichment, Microarray, Gene Expression, Stable Transfection

Fig. 6 DACH1 facilitates the recruitment of, and co-accumulates with, Ku70/Ku80 proteins at sites of DNA damage. A Co-accumulation of Ku-70/Ku-80 at laser micro irradiation-induced DSBs sites in Dach1+/+ 3T3 cells. B, C 24 h after transfection, the accumulation of DACH1 and Ku70/Ku80 in Dach1−/−3T3 cells transfected with EGFP or EGFP-tagged DACH1 and red ﬂuorescent protein (RFP)-tagged Ku70 or RFP-tagged Ku80 expression vectors were treated with laser micro-irradiation (403 nm) to induce DSBs. Time is shown after micro-irradiation. Accumulation of the transfected proteins was indicated by EGFP (green) or RFP (red) ﬂuorescence at laser-irradiated sites. Co-accumulation was visualized in yellow merged images. Time is shown in minutes and -fold increase in foci intensity is shown as mean ± SEM for N = 5 separate cells.

Figure Legend Snippet: Fig. 6 DACH1 facilitates the recruitment of, and co-accumulates with, Ku70/Ku80 proteins at sites of DNA damage. A Co-accumulation of Ku-70/Ku-80 at laser micro irradiation-induced DSBs sites in Dach1+/+ 3T3 cells. B, C 24 h after transfection, the accumulation of DACH1 and Ku70/Ku80 in Dach1−/−3T3 cells transfected with EGFP or EGFP-tagged DACH1 and red ﬂuorescent protein (RFP)-tagged Ku70 or RFP-tagged Ku80 expression vectors were treated with laser micro-irradiation (403 nm) to induce DSBs. Time is shown after micro-irradiation. Accumulation of the transfected proteins was indicated by EGFP (green) or RFP (red) ﬂuorescence at laser-irradiated sites. Co-accumulation was visualized in yellow merged images. Time is shown in minutes and -fold increase in foci intensity is shown as mean ± SEM for N = 5 separate cells.

Techniques Used: Irradiation, Transfection, Expressing

Image Search Results

Journal: Oncogene

Article Title: The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses.

doi: 10.1038/s41388-023-02668-9

Figure Lengend Snippet: Fig. 2 DACH1 deletion PCa enhances AR signaling. A Interrogation of human PCa gene expression data [26], showing candidate genetic drivers ERG, ETV1/ETV4/FLI1, SPOP, FOXA1, and unknown. Samples with DACH1 homozygous (deep) genetic deletions (29/333) are shown as an additional subtype. The AR score (the average of the AR target gene expression) refers to a group of AR-responsive genes [26], and together with the expression Z-score of the AR target genes, are shown as colorimetric scales. The AR score-based gene names are shown. The androgen receptor (AR) activity, inferred by the induction of AR target genes, was increased in DACH1 homozygous (‘deep’) deletion PCa compared with normal (P = 2 × 10−5 by t-test) and ERG mutation groups (P = 0.003 by t-test). B AR mRNA and AR protein levels, shown for each DACH1 deletion sample, were not signiﬁcantly different. C The iCluster [29], mRNA cluster, and SCNA (somatic copy-number alteration), and DNA methylation status are shown for the PCa classiﬁed by the corresponding gene deletion subtypes. D DACH1 homozygous deletions were enriched for iCluster 2 and 3 [29], mRNA cluster 2 (P = 0.0003 by Fisher exact test, SCNA (“more” somatic copy-number alteration, P = 0.0004 by Fisher exact test), but not for DNA methylation.

Article Snippet: For detection of DACH1 protein, antigen retrieval was done in Tris/EDTA buffer at pH 9 for 30min at 97 °C, followed by 30min incubation with rabbit polyclonal DACH1 antibody (Cat. #10914-1-AP, Proteintech, Rosemont, IL; dilution 1:1,000) [33], HRP-conjugated polymer (Envision FLEX, Cat#GV80011-2, Agilent), and DAB chromogen deposition.

Techniques: Gene Expression, Targeted Gene Expression, Expressing, Activity Assay, Mutagenesis, DNA Methylation Assay

Journal: Oncogene

Article Title: The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses.

doi: 10.1038/s41388-023-02668-9

Figure Lengend Snippet: Fig. 3 Prostate-speciﬁc Dach1 gene deletion promotes prostate hyperplasia and dysplasia in OncoMice (15 weeks). A Schematic representation of transgenes integrated into mice. B Representative immunohistochemistry for Dach1, with data quantitated as mean ± standard error of the mean (SEM) for N = 20 (4 separate mice, with 5 views per mouse, in each group). C Blinded quantitative histology grading of prostate of multigenic mice at 15 weeks. Data are shown as mean ± SEM for N = 15 (5 separate mice, with 3 prostate areas [anterior, ventral, lateral] per mouse) in each group). H&E staining demonstrates the presence of a focal atypical intraductal proliferation in Dach1−/−prostate, compatible with prostatic intraepithelial neoplasia (PIN). Representative immunohistochemistry with results shown as mean ± SEM for Ki-67 (n = 20, 4 separate mice for each genotype, 5 views per mouse) (D), Beclin 1 (n = 9, 3 separate mice for each genotype, 3 views per mouse) (E); and AR (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 12 for Dach1ﬂ/ﬂmice, 3 separate mice, 2 views for one mouse and 5 views for other two mice) (F). Scale bars, 50 μm. A Student’s t test was performed for all comparisons.

Techniques: Immunohistochemistry, Staining

Journal: Oncogene

Article Title: The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses.

doi: 10.1038/s41388-023-02668-9

Figure Lengend Snippet: Fig. 4 Prostate-speciﬁc Dach1 gene deletion in TRAMP mice induces PIN lesions with increased TGFβ activity. Genome-wide expression analysis of TRAMP Dach1+/+ vs. Dach1−/−PIN lesions was analyzed for enrichment of known targets of upstream regulators using Ingenuity Pathway Analysis (IPA) and represented as (A) barplot was calculated by IPA activation Z-score labeled and as (B) bubble plot with size of the bubbles proportional to –log10 p values. C IHC was conducted for SMAD activation using SMAD2P, quantitated and shown as (D) mean ± SEM (n = 15 for Dach1wt/wt mice, 3 separate mice, 5 views per mouse) (n = 10 for Dach1ﬂ/ﬂmice, 2 separate mice, 5 views per mouse). E–G Western blot of either PCa cell lines for the presence of DACH1 (E, F) or (G) TGFβ-treated (10 ng/ml for 24 h) PC3 cells illustrating induction of nuclear vimentin and cytoplasmic cyclin D1. Protein loading controls are β-tubulin (a marker of cytoplasmic proteins) and Lamin B1 (a marker for nuclear protein enrichment). H Microarray-based gene expression analysis of PC3 cells stably expressing DACH1, showing restraint of genes mediating TGFβ signaling (shown with blue arrows), including reduction of TGFB2 and TGFBR2 [33].

Techniques: Activity Assay, Genome Wide, Expressing, Activation Assay, Labeling, Western Blot, Marker, Protein Enrichment, Microarray, Gene Expression, Stable Transfection

Journal: Oncogene

Article Title: The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses.

doi: 10.1038/s41388-023-02668-9

Figure Lengend Snippet: Fig. 6 DACH1 facilitates the recruitment of, and co-accumulates with, Ku70/Ku80 proteins at sites of DNA damage. A Co-accumulation of Ku-70/Ku-80 at laser micro irradiation-induced DSBs sites in Dach1+/+ 3T3 cells. B, C 24 h after transfection, the accumulation of DACH1 and Ku70/Ku80 in Dach1−/−3T3 cells transfected with EGFP or EGFP-tagged DACH1 and red ﬂuorescent protein (RFP)-tagged Ku70 or RFP-tagged Ku80 expression vectors were treated with laser micro-irradiation (403 nm) to induce DSBs. Time is shown after micro-irradiation. Accumulation of the transfected proteins was indicated by EGFP (green) or RFP (red) ﬂuorescence at laser-irradiated sites. Co-accumulation was visualized in yellow merged images. Time is shown in minutes and -fold increase in foci intensity is shown as mean ± SEM for N = 5 separate cells.

Techniques: Irradiation, Transfection, Expressing

( A ) PAS staining in normal kidney. ( B ) Sequential sections with A and showing positive staining for DACH1 in human normal kidney. ( C ) Immunofluorescence for DACH1, showing the intensity of expression of DACH1 in glomerular is higher than tubules. ( D ) DACH1 staining in renal tissue with fibrosis, seeing renal interstitium was positive for DACH1. Original magnification × 400, Bar, 50 μm.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: ( A ) PAS staining in normal kidney. ( B ) Sequential sections with A and showing positive staining for DACH1 in human normal kidney. ( C ) Immunofluorescence for DACH1, showing the intensity of expression of DACH1 in glomerular is higher than tubules. ( D ) DACH1 staining in renal tissue with fibrosis, seeing renal interstitium was positive for DACH1. Original magnification × 400, Bar, 50 μm.

Article Snippet: DACH1 rabbit polyclonal antibody (1:1000 dilution, Proteintech, Wuhan, China), Cyclin D1, PCNA mouse monoclonal antibody (1:1000 dilution, Santa Cruz Biotechnology, Dallas, TX), Cyclin A, Cyclin B1, p21, p53 rabbit polyclonal antibody (1:500, Affinity Biosciences, USA) were used as primary antibodies.

Techniques: Staining, Immunofluorescence, Expressing

DACH1 immunostaining level in glomerulus (glom) and renal tubules (rb) in both diseased and normal samples

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: DACH1 immunostaining level in glomerulus (glom) and renal tubules (rb) in both diseased and normal samples

Techniques: Immunostaining

( A ) DACH1 staining in normal renal tissue obtained from nephrectomy specimens. ( B , C , and D ) DACH1 staining for minimal change disease, immunoglobulin A nephropathy, and idiopathic membranous nephropathy, respectively. ( E ) DACH1 expression in normal kidney tubules. ( F ) DACH1 staining in lesioned renal tissue. The DACH1 expression in glomerulus shows a positive correlation ( G ) with eGFR and a negative correlation ( H ) with serum creatinine. Such level reveals no relevance with 24 h urinary protein ( I ) and patients’ age. All photos shown display immunoperoxidase stains with hematoxylin counterstain. All tissue sections are counter-stained with periodic acid-Schiff; original magnification × 400, Bar, 50 μm.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: ( A ) DACH1 staining in normal renal tissue obtained from nephrectomy specimens. ( B , C , and D ) DACH1 staining for minimal change disease, immunoglobulin A nephropathy, and idiopathic membranous nephropathy, respectively. ( E ) DACH1 expression in normal kidney tubules. ( F ) DACH1 staining in lesioned renal tissue. The DACH1 expression in glomerulus shows a positive correlation ( G ) with eGFR and a negative correlation ( H ) with serum creatinine. Such level reveals no relevance with 24 h urinary protein ( I ) and patients’ age. All photos shown display immunoperoxidase stains with hematoxylin counterstain. All tissue sections are counter-stained with periodic acid-Schiff; original magnification × 400, Bar, 50 μm.

Techniques: Staining, Expressing

Compared with normal kidneys, DACH1 expression was decreased in kidneys associated with IgAN, MN, and MCD, ** P < 0.001, *** P < 0.01, ** P < 0.01, respectively. IgAN vs MN, IgAN vs MCD, MN vs MCD, P > 0.05, respectively, Tukey’s Multiple comparison test. The horizontal bars represent medians.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: Compared with normal kidneys, DACH1 expression was decreased in kidneys associated with IgAN, MN, and MCD, ** P < 0.001, *** P < 0.01, ** P < 0.01, respectively. IgAN vs MN, IgAN vs MCD, MN vs MCD, P > 0.05, respectively, Tukey’s Multiple comparison test. The horizontal bars represent medians.

Techniques: Expressing, Comparison

( A and C ) positive and negative control (NC) stained for DACH1 in a focal proliferative IgAN. ( B ) A modicum glomerular staining for DACH1 in scarred glomerulus in sclerotic class IgAN. ( D ) Glomerular DACH1 expression correlates strongly with eGFR in IgAN. ( E ) Glomerular DACH1 expression correlates negatively with serum creatinine in IgAN. ( F ) DACH1 detection is higher in patients with disease duration of less than 6 months. ( G ) section with positive staining for DACH1 in a phase II IMN. ( H ) Glomerular DACH1 expression correlates strongly with eGFR in IMN. ( I ) Glomerular DACH1 expression correlates negatively with 24 h urinary protein in IMN. All photos shown contain immunoperoxidase stains with hematoxylin counterstain, 400 magnification. GCS, glomerular cross section, Bar, 50 μm.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: ( A and C ) positive and negative control (NC) stained for DACH1 in a focal proliferative IgAN. ( B ) A modicum glomerular staining for DACH1 in scarred glomerulus in sclerotic class IgAN. ( D ) Glomerular DACH1 expression correlates strongly with eGFR in IgAN. ( E ) Glomerular DACH1 expression correlates negatively with serum creatinine in IgAN. ( F ) DACH1 detection is higher in patients with disease duration of less than 6 months. ( G ) section with positive staining for DACH1 in a phase II IMN. ( H ) Glomerular DACH1 expression correlates strongly with eGFR in IMN. ( I ) Glomerular DACH1 expression correlates negatively with 24 h urinary protein in IMN. All photos shown contain immunoperoxidase stains with hematoxylin counterstain, 400 magnification. GCS, glomerular cross section, Bar, 50 μm.

Techniques: Negative Control, Staining, Expressing

A representative micrograph of double immunofluorescence was performed for DACH1 (red) ( B , E , H ), Bcl-2 (green) ( A ), Bax(green) ( D ), PCNA (green) ( G ), Merged images ( C , E , I ). Original magnification is × 400, Bar, 50 μm.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: A representative micrograph of double immunofluorescence was performed for DACH1 (red) ( B , E , H ), Bcl-2 (green) ( A ), Bax(green) ( D ), PCNA (green) ( G ), Merged images ( C , E , I ). Original magnification is × 400, Bar, 50 μm.

Techniques: Immunofluorescence

A representative micrograph of double immunofluorescence was performed for DACH1(red) ( B , E , H , K ), P21 (green) ( A ), P53(green) ( D ), Cyclin A (green) ( G ), Cyclin D1 (green) ( J ), Merge images (yellow) ( C , F , I , L ). Original magnification is ×400, Bar, 50 μm.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: A representative micrograph of double immunofluorescence was performed for DACH1(red) ( B , E , H , K ), P21 (green) ( A ), P53(green) ( D ), Cyclin A (green) ( G ), Cyclin D1 (green) ( J ), Merge images (yellow) ( C , F , I , L ). Original magnification is ×400, Bar, 50 μm.

Techniques: Immunofluorescence

( A ) HK2, podocyte cultured in a permissive temperature (33°C) and cultured in a nonpermissive temperature (37°C). transfection with Plasmid.GFP. ( B ) Western blotting analysis of the DACH1 protein in Plasmid. DACH1-transfected HK2 and summarized data showing that DACH1 expression. ( C ) Western blotting analysis of the DACH1 protein in Plasmid. DACH1-transfected podocytes and summarized data showing that DACH1 expression. The data are displayed as mean ± SEM; * P < 0.05; ** P < 0.01 vs. Null-HK2 or Null-podocyte. All experiment were performed at least thrice with samples from independent experiments.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: ( A ) HK2, podocyte cultured in a permissive temperature (33°C) and cultured in a nonpermissive temperature (37°C). transfection with Plasmid.GFP. ( B ) Western blotting analysis of the DACH1 protein in Plasmid. DACH1-transfected HK2 and summarized data showing that DACH1 expression. ( C ) Western blotting analysis of the DACH1 protein in Plasmid. DACH1-transfected podocytes and summarized data showing that DACH1 expression. The data are displayed as mean ± SEM; * P < 0.05; ** P < 0.01 vs. Null-HK2 or Null-podocyte. All experiment were performed at least thrice with samples from independent experiments.

Techniques: Cell Culture, Transfection, Plasmid Preparation, Western Blot, Expressing

( A ) Representative Western blot images and ( B ) summarized data showing DACH1, p21, p53, CyclinD1, CyclinA expression with Plasmid. DACH1-transfected HK2. ( C ) representative Western blot images and ( E ) summarized data showing that the abundance DACH1, p21, p53 were increased in Plasmid. DACH1-transfected podocytes cultured in a permissive temperature (33°C). ( D ) representative Western blot images and ( F ) summarized data showing that the abundance p21, p53 were increased, while CyclinD1 was decreased in Plasmid. DACH1-transfected podocytes cultured in a permissive temperature (37°C). * P < 0.05 vs. Null-HK2 or Null-podocyte. All experiment were performed at least thrice with samples from independent experiments.

Journal: Oncotarget

Article Title: Decreased DACH1 expression in glomerulopathy is associated with disease progression and severity

doi: 10.18632/oncotarget.13470

Figure Lengend Snippet: ( A ) Representative Western blot images and ( B ) summarized data showing DACH1, p21, p53, CyclinD1, CyclinA expression with Plasmid. DACH1-transfected HK2. ( C ) representative Western blot images and ( E ) summarized data showing that the abundance DACH1, p21, p53 were increased in Plasmid. DACH1-transfected podocytes cultured in a permissive temperature (33°C). ( D ) representative Western blot images and ( F ) summarized data showing that the abundance p21, p53 were increased, while CyclinD1 was decreased in Plasmid. DACH1-transfected podocytes cultured in a permissive temperature (37°C). * P < 0.05 vs. Null-HK2 or Null-podocyte. All experiment were performed at least thrice with samples from independent experiments.

Techniques: Western Blot, Expressing, Plasmid Preparation, Transfection, Cell Culture

Fig. 1. The expression pattern of Dach1 message in the developing limb bud changed with the stage of mesenchymal cell differentiation. Mouse forelimb buds and whole embryos were stained to reveal Dach1 expression by whole-mount in situ hybridization. A: At 10 days postcoitus (dpc), Dach1 was concentrated in four regions of the limb bud (1–4), with region 2 overlapping with the zone of polarizing activity. B: By 11 dpc, Dach1 expression was predominantly in the subridge mesenchyme and was excluded from the central area of precartilaginous cell condensation. C: At 12 dpc, Dach1 was expressed at the tips of each digit, capped the digital cartilages distally, and was present in the interdigital web space. D: The limb bud was a prominent site of Dach1 expression in the 12 dpc embryo along with the forebrain, eye, genital eminence, and caudal neural tube. Development of the hindlimb is delayed in comparison to the forelimb, and the staining pattern of the hindlimb appeared intermediate between that of the 11 and 12 dpc forelimb.

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Fibroblast growth factor signaling regulates Dach1 expression during skeletal development.

doi: 10.1002/dvdy.10132

Figure Lengend Snippet: Fig. 1. The expression pattern of Dach1 message in the developing limb bud changed with the stage of mesenchymal cell differentiation. Mouse forelimb buds and whole embryos were stained to reveal Dach1 expression by whole-mount in situ hybridization. A: At 10 days postcoitus (dpc), Dach1 was concentrated in four regions of the limb bud (1–4), with region 2 overlapping with the zone of polarizing activity. B: By 11 dpc, Dach1 expression was predominantly in the subridge mesenchyme and was excluded from the central area of precartilaginous cell condensation. C: At 12 dpc, Dach1 was expressed at the tips of each digit, capped the digital cartilages distally, and was present in the interdigital web space. D: The limb bud was a prominent site of Dach1 expression in the 12 dpc embryo along with the forebrain, eye, genital eminence, and caudal neural tube. Development of the hindlimb is delayed in comparison to the forelimb, and the staining pattern of the hindlimb appeared intermediate between that of the 11 and 12 dpc forelimb.

Article Snippet: Cell Culture and Purification of Murine Recombinant Dach1 A recombinant adenovirus for the expression of Dach1 was constructed (Qbiogene, Carlsbad, CA) by using the full-length, FLAG-tagged cDNA from pDacCF (Ayres et al., 2001).

Techniques: Expressing, Cell Differentiation, Staining, In Situ Hybridization, Activity Assay, Comparison

Fig. 2. Dach1 was localized to proliferating mesenchymal cells of the limb buds by immunohistochemistry. Immunoreactive product is shown in brown. A: Staining with afﬁnity puriﬁed anti-Dach1 antibodies revealed labeling of mesenchymal cells in the 10 days postcoitus (dpc) forelimb bud, corresponding to the anterior and posterior areas of highest Dach1 message accumulation (Fig. 1A). B: At 12 dpc, the Dach1 protein was concentrated at the distal tips of the developing digits (arrow) and sur- rounding the distal ends of the digital cartilages (open arrowhead). There was relatively intense staining of the mesenchyme anterior to the ﬁrst digit and posterior to the last digit (arrowheads). C: These areas of Dach1 staining corresponded to areas stained for proliferating cell nuclear anti- gen (PCNA). D: Double immunoﬂuorescent staining for Dach1 (in red) and PCNA (in green) in the subectodermal mesenchyme anterior to the ﬁrst digit revealed many nuclei labeled for both antigens (in yellow). Scale bar 25 m in D.

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Fibroblast growth factor signaling regulates Dach1 expression during skeletal development.

doi: 10.1002/dvdy.10132

Figure Lengend Snippet: Fig. 2. Dach1 was localized to proliferating mesenchymal cells of the limb buds by immunohistochemistry. Immunoreactive product is shown in brown. A: Staining with afﬁnity puriﬁed anti-Dach1 antibodies revealed labeling of mesenchymal cells in the 10 days postcoitus (dpc) forelimb bud, corresponding to the anterior and posterior areas of highest Dach1 message accumulation (Fig. 1A). B: At 12 dpc, the Dach1 protein was concentrated at the distal tips of the developing digits (arrow) and sur- rounding the distal ends of the digital cartilages (open arrowhead). There was relatively intense staining of the mesenchyme anterior to the ﬁrst digit and posterior to the last digit (arrowheads). C: These areas of Dach1 staining corresponded to areas stained for proliferating cell nuclear anti- gen (PCNA). D: Double immunoﬂuorescent staining for Dach1 (in red) and PCNA (in green) in the subectodermal mesenchyme anterior to the ﬁrst digit revealed many nuclei labeled for both antigens (in yellow). Scale bar 25 m in D.

Techniques: Immunohistochemistry, Staining, Labeling

Fig. 3. Dach1 was expressed in the 17 days postcoitus (dpc) mouse hindlimb. A: Dach1 immunostaining labeled chondrocytes of the phalan- ges (arrows) and dermal cells of the foot pad (arrowheads). B: At the distal tips of the digits, Dach1 was stained in the dermis (arrowheads) and in the chondrocytes of the distal phalanges (arrow). C: Staining of the proximal phalanges revealed more mature chondrocytes at the center of the anlagen (arrow) and chondrocytes of the developing joint (arrow- heads). D: Incubating the anti-Dach1 antiserum with puriﬁed Dach1 protein before staining blocked the activity of this antiserum and con- ﬁrmed its speciﬁcity. Scale bars 100 m in A–D.

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Fibroblast growth factor signaling regulates Dach1 expression during skeletal development.

doi: 10.1002/dvdy.10132

Figure Lengend Snippet: Fig. 3. Dach1 was expressed in the 17 days postcoitus (dpc) mouse hindlimb. A: Dach1 immunostaining labeled chondrocytes of the phalan- ges (arrows) and dermal cells of the foot pad (arrowheads). B: At the distal tips of the digits, Dach1 was stained in the dermis (arrowheads) and in the chondrocytes of the distal phalanges (arrow). C: Staining of the proximal phalanges revealed more mature chondrocytes at the center of the anlagen (arrow) and chondrocytes of the developing joint (arrow- heads). D: Incubating the anti-Dach1 antiserum with puriﬁed Dach1 protein before staining blocked the activity of this antiserum and con- ﬁrmed its speciﬁcity. Scale bars 100 m in A–D.

Techniques: Immunostaining, Labeling, Staining, Activity Assay

Fig. 8. FGF4 and FGF8 can substitute for the apical ectodermal ridge (AER) in maintaining Dach1 expression. (A) The expression of Dach1 in the forelimb bud dissected at 11.5 days postcoitus (dpc) and cultured for 24 hr is broader than that dissected at 11 dpc (Fig. 7A). B: When the AER was removed from the limb bud before culture, Dach1 expression was down-regulated. When the AER was removed and beads soaked in 1 mg/ml FGF4 (C) or FGF8 (D) were implanted into the periphery of the bud, Dach1 expression was maintained.

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Fibroblast growth factor signaling regulates Dach1 expression during skeletal development.

doi: 10.1002/dvdy.10132

Figure Lengend Snippet: Fig. 8. FGF4 and FGF8 can substitute for the apical ectodermal ridge (AER) in maintaining Dach1 expression. (A) The expression of Dach1 in the forelimb bud dissected at 11.5 days postcoitus (dpc) and cultured for 24 hr is broader than that dissected at 11 dpc (Fig. 7A). B: When the AER was removed from the limb bud before culture, Dach1 expression was down-regulated. When the AER was removed and beads soaked in 1 mg/ml FGF4 (C) or FGF8 (D) were implanted into the periphery of the bud, Dach1 expression was maintained.

Techniques: Expressing, Cell Culture

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