Review



mouse anti dzip1l  (Proteintech)


Bioz Verified Symbol Proteintech is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    Proteintech mouse anti dzip1l
    Mapping of a new ARPKD locus on 3q22.1-q23 and identification of <t>DZIP1L</t> mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).
    Mouse Anti Dzip1l, supplied by Proteintech, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+dzip1l/DZIP1+Antibody/pmc05687889-566-38-49
    Average 88 stars, based on 3 article reviews
    mouse anti dzip1l - by Bioz Stars, 2026-09
    88/100 stars

    Images

    1) Product Images from "Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease"

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    Journal: Nature genetics

    doi: 10.1038/ng.3871

    Mapping of a new ARPKD locus on 3q22.1-q23 and identification of DZIP1L mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).
    Figure Legend Snippet: Mapping of a new ARPKD locus on 3q22.1-q23 and identification of DZIP1L mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).

    Techniques Used: Genome Wide, Multiplex Assay, Sequencing, Mutagenesis

    Mutations of DZIP1L in patients with ARPKD.
    Figure Legend Snippet: Mutations of DZIP1L in patients with ARPKD.

    Techniques Used: Mutagenesis

    DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.
    Figure Legend Snippet: DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.

    Techniques Used: Staining, Microscopy, Labeling

    Dzip1l is required for ciliary differentiation in the zebrafish kidney tubule. (a) Primary cilia (short arrows; myotome) and mono motile cilia (long arrows; pronephric duct) in a 24 hpf wild-type embryo. (b) No obvious difference in primary (short arrows) and mono motile cilia (long arrows) differentiation in a dzip1l splice morpholino (e5i5) injected embryo at 24 hpf (n=10). (c) MCC motile cilia bundles (arrows) in the pronephric duct of a wild-type embryo at 48 hpf (arrows). (d) MCC motile cilia bundles (arrows) are reduced in a 48 hpf maternal-zygotic dzip1l mutant zebrafish embryo (n=8). (e) Quantification of MCC cilia bundles in wild-type and maternal-zygotic dzip1l mutant zebrafish embryos. 8 embryos from each group were analyzed. **p=0.0017 based on unpaired Student’s t-test. Error bars show SEM. In all panels, cilia were stained with anti-acetylated tubulin antibody (magenta) and nuclei with DAPI (blue); in panels a,b,f-i cell membranes were stained with β-catenin antibodies (green). Scale bars in a,b,f-i = 10µm; c,d = 100µm.
    Figure Legend Snippet: Dzip1l is required for ciliary differentiation in the zebrafish kidney tubule. (a) Primary cilia (short arrows; myotome) and mono motile cilia (long arrows; pronephric duct) in a 24 hpf wild-type embryo. (b) No obvious difference in primary (short arrows) and mono motile cilia (long arrows) differentiation in a dzip1l splice morpholino (e5i5) injected embryo at 24 hpf (n=10). (c) MCC motile cilia bundles (arrows) in the pronephric duct of a wild-type embryo at 48 hpf (arrows). (d) MCC motile cilia bundles (arrows) are reduced in a 48 hpf maternal-zygotic dzip1l mutant zebrafish embryo (n=8). (e) Quantification of MCC cilia bundles in wild-type and maternal-zygotic dzip1l mutant zebrafish embryos. 8 embryos from each group were analyzed. **p=0.0017 based on unpaired Student’s t-test. Error bars show SEM. In all panels, cilia were stained with anti-acetylated tubulin antibody (magenta) and nuclei with DAPI (blue); in panels a,b,f-i cell membranes were stained with β-catenin antibodies (green). Scale bars in a,b,f-i = 10µm; c,d = 100µm.

    Techniques Used: Injection, Mutagenesis, Staining

    DZIP1L associates with the ciliary transition zone protein SEPT2. (a) Schematic diagram of SEPT2, DZIP1L and a series of DZIP1L deletion mutants. (b) Interaction between SEPT2 (N-terminal FLAG) and DZIP1L and various DZIP1L deletion mutants (C-terminal HA) as determined by IP following transfection in HEK293T cells. (c,d) Endogenous interaction between DZIP1L and SEPT2 in RPE-1 cells. IP was performed with either the anti-DZIP1L antibody (Abnova; c) or the anti-SEPT2 antibody (d). (e) Expression of DZIP1L and SEPT2 in total cell lysate. (f) Co-localization of DZIP1L (cyan) and SEPT2 (magenta) in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary axoneme (green). (g,h) SEPT2 (green) localization to the transition zone was unaffected in DZIP1L mutant dermal fibroblasts from individual B155 (p.Gln155*). Ciliary axonemes and basal bodies were labelled with anti-acetylated tubulin and anti-γ-tubulin antibodies (magenta), respectively. Scale bars in f-h = 1µm. All co-IP experiments were repeated at least three times.
    Figure Legend Snippet: DZIP1L associates with the ciliary transition zone protein SEPT2. (a) Schematic diagram of SEPT2, DZIP1L and a series of DZIP1L deletion mutants. (b) Interaction between SEPT2 (N-terminal FLAG) and DZIP1L and various DZIP1L deletion mutants (C-terminal HA) as determined by IP following transfection in HEK293T cells. (c,d) Endogenous interaction between DZIP1L and SEPT2 in RPE-1 cells. IP was performed with either the anti-DZIP1L antibody (Abnova; c) or the anti-SEPT2 antibody (d). (e) Expression of DZIP1L and SEPT2 in total cell lysate. (f) Co-localization of DZIP1L (cyan) and SEPT2 (magenta) in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary axoneme (green). (g,h) SEPT2 (green) localization to the transition zone was unaffected in DZIP1L mutant dermal fibroblasts from individual B155 (p.Gln155*). Ciliary axonemes and basal bodies were labelled with anti-acetylated tubulin and anti-γ-tubulin antibodies (magenta), respectively. Scale bars in f-h = 1µm. All co-IP experiments were repeated at least three times.

    Techniques Used: Transfection, Expressing, Mutagenesis, Co-Immunoprecipitation Assay

    Loss of DZIP1L affects the localization of PC1 and PC2 to the ciliary membrane. (a) In most wild-type ciliated MEFs PC1 (green; 7e12 mAb, Abcam) staining was distributed along the length of the axoneme as marked with ARL13B (magenta). (b) By contrast PC1 staining was more often concentrated at the proximal end of the axoneme in Dzip1lwpy/wpy MEFs (c) Quantification of the percentage of ciliated cells with PC1 along the axoneme (n=6, derived from 3 MEF cell lines from independent embryos, each counted in two separate experiments; between 50–185 cells counted per replicate, total 737 Dzip1l+/+ and 649 Dzip1lwpy/wpy cells counted). (d,e) Decreased PC1 (green) localization in the ciliary membrane in human dermal fibroblasts from affected individual B155 (p.Gln155*). ARL13B (magenta) marks the ciliary axoneme. (g,h) Decreased PC2 localization (green) in the ciliary membrane in DZIP1L mutant human dermal fibroblasts. Acetylated-α–tubulin and γ-tubulin (magenta) mark the cilia. (f,i) Quantification of the percentage of ciliated cells with PC1 and PC2 along the axoneme, respectively, in control and DZIP1L mutant human dermal fibroblasts. For human cells, cilia were counted in three experiments, with cells from three independent coverslips counted for each experiment (approximately 100 ciliated cells counted on each coverslip). In all cases, quantification included cilia with or without detectable PC1 or PC2 staining. Statistical analyses based on unpaired Student’s t-test. Error bars show SEM. For PC1 staining on MEFs, ****p<0.0001; PC1 and PC2 staining on human dermal fibroblasts, ***p<0.001. Scale bars in a,b = 2µm; d,e,g,h = 1µm.
    Figure Legend Snippet: Loss of DZIP1L affects the localization of PC1 and PC2 to the ciliary membrane. (a) In most wild-type ciliated MEFs PC1 (green; 7e12 mAb, Abcam) staining was distributed along the length of the axoneme as marked with ARL13B (magenta). (b) By contrast PC1 staining was more often concentrated at the proximal end of the axoneme in Dzip1lwpy/wpy MEFs (c) Quantification of the percentage of ciliated cells with PC1 along the axoneme (n=6, derived from 3 MEF cell lines from independent embryos, each counted in two separate experiments; between 50–185 cells counted per replicate, total 737 Dzip1l+/+ and 649 Dzip1lwpy/wpy cells counted). (d,e) Decreased PC1 (green) localization in the ciliary membrane in human dermal fibroblasts from affected individual B155 (p.Gln155*). ARL13B (magenta) marks the ciliary axoneme. (g,h) Decreased PC2 localization (green) in the ciliary membrane in DZIP1L mutant human dermal fibroblasts. Acetylated-α–tubulin and γ-tubulin (magenta) mark the cilia. (f,i) Quantification of the percentage of ciliated cells with PC1 and PC2 along the axoneme, respectively, in control and DZIP1L mutant human dermal fibroblasts. For human cells, cilia were counted in three experiments, with cells from three independent coverslips counted for each experiment (approximately 100 ciliated cells counted on each coverslip). In all cases, quantification included cilia with or without detectable PC1 or PC2 staining. Statistical analyses based on unpaired Student’s t-test. Error bars show SEM. For PC1 staining on MEFs, ****p<0.0001; PC1 and PC2 staining on human dermal fibroblasts, ***p<0.001. Scale bars in a,b = 2µm; d,e,g,h = 1µm.

    Techniques Used: Staining, Derivative Assay, Mutagenesis



    Similar Products

    mouse polyclonal anti-human dzip1l protein antibody  (Abnova)
    90
    Abnova mouse polyclonal anti-human dzip1l protein antibody
    Mouse Polyclonal Anti Human Dzip1l Protein Antibody, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+dzip1l/anti+dzip1l+antibody/pm35211789-85-8-16
    Average 90 stars, based on 1 article reviews
    mouse polyclonal anti-human dzip1l protein antibody - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    mouse anti dzip1l  (Proteintech)
    88
    Proteintech mouse anti dzip1l
    Mapping of a new ARPKD locus on 3q22.1-q23 and identification of <t>DZIP1L</t> mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).
    Mouse Anti Dzip1l, supplied by Proteintech, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+dzip1l/DZIP1+Antibody/pmc05687889-566-38-49
    Average 88 stars, based on 1 article reviews
    mouse anti dzip1l - by Bioz Stars, 2026-09
    88/100 stars
    		  Buy from Supplier

    Image Search Results


    Mapping of a new ARPKD locus on 3q22.1-q23 and identification of DZIP1L mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: Mapping of a new ARPKD locus on 3q22.1-q23 and identification of DZIP1L mutations. Genome-wide SNP analysis performed in two unrelated consanguineous multiplex pedigrees (shown at top) with a total of five children affected by ARPKD, resulted in identification of a single overlapping 7.5 Mb region of homozygosity on chromosome 3q22.1-q23. By various sequencing approaches, we identified different homozygous DZIP1L mutations in these and other consanguineous families with ARPKD (see text for details). In parallel, we identified an ENU-induced recessive mouse model (see Fig. 2) bearing the homozygous Dzip1l nonsense mutation c.1123C>T (p.Gln375*) (in blue).

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Genome Wide, Multiplex Assay, Sequencing, Mutagenesis

    Mutations of DZIP1L in patients with ARPKD.

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: Mutations of DZIP1L in patients with ARPKD.

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Mutagenesis

    DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Staining, Microscopy, Labeling

    Dzip1l is required for ciliary differentiation in the zebrafish kidney tubule. (a) Primary cilia (short arrows; myotome) and mono motile cilia (long arrows; pronephric duct) in a 24 hpf wild-type embryo. (b) No obvious difference in primary (short arrows) and mono motile cilia (long arrows) differentiation in a dzip1l splice morpholino (e5i5) injected embryo at 24 hpf (n=10). (c) MCC motile cilia bundles (arrows) in the pronephric duct of a wild-type embryo at 48 hpf (arrows). (d) MCC motile cilia bundles (arrows) are reduced in a 48 hpf maternal-zygotic dzip1l mutant zebrafish embryo (n=8). (e) Quantification of MCC cilia bundles in wild-type and maternal-zygotic dzip1l mutant zebrafish embryos. 8 embryos from each group were analyzed. **p=0.0017 based on unpaired Student’s t-test. Error bars show SEM. In all panels, cilia were stained with anti-acetylated tubulin antibody (magenta) and nuclei with DAPI (blue); in panels a,b,f-i cell membranes were stained with β-catenin antibodies (green). Scale bars in a,b,f-i = 10µm; c,d = 100µm.

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: Dzip1l is required for ciliary differentiation in the zebrafish kidney tubule. (a) Primary cilia (short arrows; myotome) and mono motile cilia (long arrows; pronephric duct) in a 24 hpf wild-type embryo. (b) No obvious difference in primary (short arrows) and mono motile cilia (long arrows) differentiation in a dzip1l splice morpholino (e5i5) injected embryo at 24 hpf (n=10). (c) MCC motile cilia bundles (arrows) in the pronephric duct of a wild-type embryo at 48 hpf (arrows). (d) MCC motile cilia bundles (arrows) are reduced in a 48 hpf maternal-zygotic dzip1l mutant zebrafish embryo (n=8). (e) Quantification of MCC cilia bundles in wild-type and maternal-zygotic dzip1l mutant zebrafish embryos. 8 embryos from each group were analyzed. **p=0.0017 based on unpaired Student’s t-test. Error bars show SEM. In all panels, cilia were stained with anti-acetylated tubulin antibody (magenta) and nuclei with DAPI (blue); in panels a,b,f-i cell membranes were stained with β-catenin antibodies (green). Scale bars in a,b,f-i = 10µm; c,d = 100µm.

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Injection, Mutagenesis, Staining

    DZIP1L associates with the ciliary transition zone protein SEPT2. (a) Schematic diagram of SEPT2, DZIP1L and a series of DZIP1L deletion mutants. (b) Interaction between SEPT2 (N-terminal FLAG) and DZIP1L and various DZIP1L deletion mutants (C-terminal HA) as determined by IP following transfection in HEK293T cells. (c,d) Endogenous interaction between DZIP1L and SEPT2 in RPE-1 cells. IP was performed with either the anti-DZIP1L antibody (Abnova; c) or the anti-SEPT2 antibody (d). (e) Expression of DZIP1L and SEPT2 in total cell lysate. (f) Co-localization of DZIP1L (cyan) and SEPT2 (magenta) in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary axoneme (green). (g,h) SEPT2 (green) localization to the transition zone was unaffected in DZIP1L mutant dermal fibroblasts from individual B155 (p.Gln155*). Ciliary axonemes and basal bodies were labelled with anti-acetylated tubulin and anti-γ-tubulin antibodies (magenta), respectively. Scale bars in f-h = 1µm. All co-IP experiments were repeated at least three times.

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: DZIP1L associates with the ciliary transition zone protein SEPT2. (a) Schematic diagram of SEPT2, DZIP1L and a series of DZIP1L deletion mutants. (b) Interaction between SEPT2 (N-terminal FLAG) and DZIP1L and various DZIP1L deletion mutants (C-terminal HA) as determined by IP following transfection in HEK293T cells. (c,d) Endogenous interaction between DZIP1L and SEPT2 in RPE-1 cells. IP was performed with either the anti-DZIP1L antibody (Abnova; c) or the anti-SEPT2 antibody (d). (e) Expression of DZIP1L and SEPT2 in total cell lysate. (f) Co-localization of DZIP1L (cyan) and SEPT2 (magenta) in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary axoneme (green). (g,h) SEPT2 (green) localization to the transition zone was unaffected in DZIP1L mutant dermal fibroblasts from individual B155 (p.Gln155*). Ciliary axonemes and basal bodies were labelled with anti-acetylated tubulin and anti-γ-tubulin antibodies (magenta), respectively. Scale bars in f-h = 1µm. All co-IP experiments were repeated at least three times.

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Transfection, Expressing, Mutagenesis, Co-Immunoprecipitation Assay

    Loss of DZIP1L affects the localization of PC1 and PC2 to the ciliary membrane. (a) In most wild-type ciliated MEFs PC1 (green; 7e12 mAb, Abcam) staining was distributed along the length of the axoneme as marked with ARL13B (magenta). (b) By contrast PC1 staining was more often concentrated at the proximal end of the axoneme in Dzip1lwpy/wpy MEFs (c) Quantification of the percentage of ciliated cells with PC1 along the axoneme (n=6, derived from 3 MEF cell lines from independent embryos, each counted in two separate experiments; between 50–185 cells counted per replicate, total 737 Dzip1l+/+ and 649 Dzip1lwpy/wpy cells counted). (d,e) Decreased PC1 (green) localization in the ciliary membrane in human dermal fibroblasts from affected individual B155 (p.Gln155*). ARL13B (magenta) marks the ciliary axoneme. (g,h) Decreased PC2 localization (green) in the ciliary membrane in DZIP1L mutant human dermal fibroblasts. Acetylated-α–tubulin and γ-tubulin (magenta) mark the cilia. (f,i) Quantification of the percentage of ciliated cells with PC1 and PC2 along the axoneme, respectively, in control and DZIP1L mutant human dermal fibroblasts. For human cells, cilia were counted in three experiments, with cells from three independent coverslips counted for each experiment (approximately 100 ciliated cells counted on each coverslip). In all cases, quantification included cilia with or without detectable PC1 or PC2 staining. Statistical analyses based on unpaired Student’s t-test. Error bars show SEM. For PC1 staining on MEFs, ****p<0.0001; PC1 and PC2 staining on human dermal fibroblasts, ***p<0.001. Scale bars in a,b = 2µm; d,e,g,h = 1µm.

    Journal: Nature genetics

    Article Title: Mutations in DZIP1L , which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    doi: 10.1038/ng.3871

    Figure Lengend Snippet: Loss of DZIP1L affects the localization of PC1 and PC2 to the ciliary membrane. (a) In most wild-type ciliated MEFs PC1 (green; 7e12 mAb, Abcam) staining was distributed along the length of the axoneme as marked with ARL13B (magenta). (b) By contrast PC1 staining was more often concentrated at the proximal end of the axoneme in Dzip1lwpy/wpy MEFs (c) Quantification of the percentage of ciliated cells with PC1 along the axoneme (n=6, derived from 3 MEF cell lines from independent embryos, each counted in two separate experiments; between 50–185 cells counted per replicate, total 737 Dzip1l+/+ and 649 Dzip1lwpy/wpy cells counted). (d,e) Decreased PC1 (green) localization in the ciliary membrane in human dermal fibroblasts from affected individual B155 (p.Gln155*). ARL13B (magenta) marks the ciliary axoneme. (g,h) Decreased PC2 localization (green) in the ciliary membrane in DZIP1L mutant human dermal fibroblasts. Acetylated-α–tubulin and γ-tubulin (magenta) mark the cilia. (f,i) Quantification of the percentage of ciliated cells with PC1 and PC2 along the axoneme, respectively, in control and DZIP1L mutant human dermal fibroblasts. For human cells, cilia were counted in three experiments, with cells from three independent coverslips counted for each experiment (approximately 100 ciliated cells counted on each coverslip). In all cases, quantification included cilia with or without detectable PC1 or PC2 staining. Statistical analyses based on unpaired Student’s t-test. Error bars show SEM. For PC1 staining on MEFs, ****p<0.0001; PC1 and PC2 staining on human dermal fibroblasts, ***p<0.001. Scale bars in a,b = 2µm; d,e,g,h = 1µm.

    Article Snippet: #17711-1-AP; 1:500 for IF); rabbit-anti-α-tubulin (Sigma-Aldrich T5192; 1:500 for IF); rabbit-anti-acetylated-α–tubulin (Cell Signalling Technology #5335; 1:800 for IF); mouse-anti-acetylated-α–tubulin (Sigma-Aldrich 6-11B-1; #T 6793; 1:500–700 for IF); mouse-anti-γ-tubulin (Sigma-Aldrich T6557; 1:500 for IF), rabbit-anti-DZIP1L (Sigma-Aldrich #HPA030404; 1:500 for IF); mouse-anti-DZIP1L (Abnova-MaxPab H00199221-B01P; 1:500 for IF; 1:1000 for western blot); rabbit-anti-DZIP1 (Proteintech Group, #13779-1-AP; 1:500 for IF); rabbit-anti-SEPT2 (Sigma-Aldrich, #HPA018481;1:2000 for western blot; 1:500 for IF); rabbit-anti-CEP164 (SAB3500022; Sigma), rabbit-anti-TCTN1 (ProteinTech Group, #15004-1-AP; 1:500 for IF); mouse-anti-Polycystin 1 (Abcam, #AB74115; Santa Cruz Biotechnology, # sc-130554; 1:200 for IF); rabbit-anti-Polycystin 2 (MV12; gift from R. Witzgall; 1:500 for IF); rabbit-anti-FPC (PKHD1) 9 (hAR-Np and hAR-c2P; gift from G. Wu; 1:10 and 1:20, respectively); rabbit-anti-SMO 51 was a kind gift of Rajat Rohatgi (Stanford University; 1:500 for IF); rabbit-anti-IFT88 (Proteintech 13967-1-AP; 1:300 for IF); anti-Aquaporin 2 antibody (Millipore; Cat # AB3274; 1:200 for IF); rabbit-anti-β-catenin antibody (Abcam, #AB6302; 1:500 for IF); goat-anti-HNF3β (M-20) (Santa-Cruz, #sc-6544; 1:50 for IF); goat-anti-SHH(N-19) (Santa-Cruz, #sc-1194; 1:100 for IF); the mouse monoclonal antibodies for Isl-1 (1:200 for IF), Pax6 (1:10 for IF) and Nkx2.2 (1:50 for IF) were obtained from the Developmental Studies Hybridoma Bank (developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.Bank); biotinylated Lotus Tetragonolobus Lectin (LTL) (Vector laboratories; Cat # B1325; 1:200).

    Techniques: Staining, Derivative Assay, Mutagenesis