90437 Search Results


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    • rabbit anti wdr60  (Novus Biologicals)
    88
    Novus Biologicals rabbit anti wdr60
    (A) Cilia stained with the markers Arl13b (green) and acetylated tubulin (AcTub, red) in RPE1 <t>WDR60</t> and WDR34 KO cell lines. Scale bars 5µ.m. (B) Percentage of ciliated cells (n=3; 656 WT, 430 WDR34 KO#1, 296 WDR34 KO#2, 397 WDR34 KO CTRL, 343 WDR60 KO and 195 WDR60 KO CTRL cells quantified). (C) Cilia length in WDR60 and WDR34 KO compared with WT cells and CRISPR control cells lines (n=3; 120 WT, 158 WDR60 KO, 138 WDR60 KO CTRL and 30 WDR34#1 cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (D-Hi) Representative 70 nm thick EM sections of (D) WT, (E) WDR34 KO and (F, G-Hi) WDR60 KO cilia. (E) Six serial sections through a WDR34 KO cilium showing no axoneme extension. (G) Two serial sections through a WDR60 KO cilium showing. Arrows point to the bulbous ciliary tip and to a membrane protrusion containing membrane vesicles; enlargements are shown to the right (H and Hi). Scale bar length and magnification is indicated on the images.
    Rabbit Anti Wdr60, supplied by Novus Biologicals, 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/result/rabbit anti wdr60/product/Novus Biologicals
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti wdr60 - by Bioz Stars, 2023-09
    88/100 stars
    		  Buy from Supplier
    rabbit polyclonal anti caspase 3 ab 90437 antibodies  (Abcam)
    86
    Abcam rabbit polyclonal anti caspase 3 ab 90437 antibodies
    Effect of baicalin on cellular apoptosis in mouse embryos. The nuclei of blastocysts (n = 30) were stained with Hoechst 33342 to examine cell apoptosis (Bar = 100 μm). Blastocysts from the in vitro (left), baicalin-treated (middle), and in vivo (right) groups showing higher degree of apoptosis of nuclei for in vitro group (white arrows) than the baicalin treated or in vivo group (A). Relative mRNA and protein expression levels <t>of</t> <t>Caspase-3</t> (B), BAX (C) and BCL-2 (D), respectively, in mouse blastocysts (n = 30) from the in vitro , baicalin treated and in vivo groups. * P < 0.05 vs . in vivo control group and ** P < 0.01 vs . in vivo control group; # P < 0.05 vs . IVC group and ## P < 0.01 vs . IVC group.
    Rabbit Polyclonal Anti Caspase 3 Ab 90437 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti caspase 3 ab 90437 antibodies/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti caspase 3 ab 90437 antibodies - by Bioz Stars, 2023-09
    86/100 stars
    		  Buy from Supplier
    model 90437  (Thermo Fisher)
    86
    Thermo Fisher model 90437
    Effect of baicalin on cellular apoptosis in mouse embryos. The nuclei of blastocysts (n = 30) were stained with Hoechst 33342 to examine cell apoptosis (Bar = 100 μm). Blastocysts from the in vitro (left), baicalin-treated (middle), and in vivo (right) groups showing higher degree of apoptosis of nuclei for in vitro group (white arrows) than the baicalin treated or in vivo group (A). Relative mRNA and protein expression levels <t>of</t> <t>Caspase-3</t> (B), BAX (C) and BCL-2 (D), respectively, in mouse blastocysts (n = 30) from the in vitro , baicalin treated and in vivo groups. * P < 0.05 vs . in vivo control group and ** P < 0.01 vs . in vivo control group; # P < 0.05 vs . IVC group and ## P < 0.01 vs . IVC group.
    Model 90437, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/model 90437/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    model 90437 - by Bioz Stars, 2023-09
    86/100 stars
    		  Buy from Supplier
    plvx cul4b 3xflag ires puro plasmid 90437  (Addgene inc)
    N/A
    Standard format Plasmid sent in bacteria as agar stab
    		   Buy from Supplier

    Image Search Results


    (A) Cilia stained with the markers Arl13b (green) and acetylated tubulin (AcTub, red) in RPE1 WDR60 and WDR34 KO cell lines. Scale bars 5µ.m. (B) Percentage of ciliated cells (n=3; 656 WT, 430 WDR34 KO#1, 296 WDR34 KO#2, 397 WDR34 KO CTRL, 343 WDR60 KO and 195 WDR60 KO CTRL cells quantified). (C) Cilia length in WDR60 and WDR34 KO compared with WT cells and CRISPR control cells lines (n=3; 120 WT, 158 WDR60 KO, 138 WDR60 KO CTRL and 30 WDR34#1 cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (D-Hi) Representative 70 nm thick EM sections of (D) WT, (E) WDR34 KO and (F, G-Hi) WDR60 KO cilia. (E) Six serial sections through a WDR34 KO cilium showing no axoneme extension. (G) Two serial sections through a WDR60 KO cilium showing. Arrows point to the bulbous ciliary tip and to a membrane protrusion containing membrane vesicles; enlargements are shown to the right (H and Hi). Scale bar length and magnification is indicated on the images.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A) Cilia stained with the markers Arl13b (green) and acetylated tubulin (AcTub, red) in RPE1 WDR60 and WDR34 KO cell lines. Scale bars 5µ.m. (B) Percentage of ciliated cells (n=3; 656 WT, 430 WDR34 KO#1, 296 WDR34 KO#2, 397 WDR34 KO CTRL, 343 WDR60 KO and 195 WDR60 KO CTRL cells quantified). (C) Cilia length in WDR60 and WDR34 KO compared with WT cells and CRISPR control cells lines (n=3; 120 WT, 158 WDR60 KO, 138 WDR60 KO CTRL and 30 WDR34#1 cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (D-Hi) Representative 70 nm thick EM sections of (D) WT, (E) WDR34 KO and (F, G-Hi) WDR60 KO cilia. (E) Six serial sections through a WDR34 KO cilium showing no axoneme extension. (G) Two serial sections through a WDR60 KO cilium showing. Arrows point to the bulbous ciliary tip and to a membrane protrusion containing membrane vesicles; enlargements are shown to the right (H and Hi). Scale bar length and magnification is indicated on the images.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Staining, CRISPR, MANN-WHITNEY

    (A, B) Localization of (A) IFT88 and (B) IFT57 in WT, WDR60 KO, and WDR34 KO#1 cells. Line graphs show lines scans of IFT intensity along the length of a representative cilium from WT (Ai and Bi, green), WDR60 KO (Aii and Bii, orange) and WDR34 KO (Aiii and Biii, yellow) cells. (C, D) Quantification of IFT-B localization within cilia in WT and WDR60 KO cells (C, IFT88 102 WT and 101 WDR60 cells quantified; D, IFT57 106 WT and 98 WDR60 KO cells quantified; n=3 independent experiments). Mann-Whitney test was used, p-value: ****=<0.0001. (E) Endogenous IFT20 accumulates at the ciliary tip in WDR60 KO cells. (F) Quantification of IFT20 localization within cilia in WDR60 KO cells (n=3 independent experiments, 102 WT and 100 WDR60 KO cells quantified). (G) Localization of IFT20-GFP in WT and WDR60 KO fixed cells. Scale bar, all panels = 5 µm. Arrows point to the ciliary base.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A, B) Localization of (A) IFT88 and (B) IFT57 in WT, WDR60 KO, and WDR34 KO#1 cells. Line graphs show lines scans of IFT intensity along the length of a representative cilium from WT (Ai and Bi, green), WDR60 KO (Aii and Bii, orange) and WDR34 KO (Aiii and Biii, yellow) cells. (C, D) Quantification of IFT-B localization within cilia in WT and WDR60 KO cells (C, IFT88 102 WT and 101 WDR60 cells quantified; D, IFT57 106 WT and 98 WDR60 KO cells quantified; n=3 independent experiments). Mann-Whitney test was used, p-value: ****=<0.0001. (E) Endogenous IFT20 accumulates at the ciliary tip in WDR60 KO cells. (F) Quantification of IFT20 localization within cilia in WDR60 KO cells (n=3 independent experiments, 102 WT and 100 WDR60 KO cells quantified). (G) Localization of IFT20-GFP in WT and WDR60 KO fixed cells. Scale bar, all panels = 5 µm. Arrows point to the ciliary base.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: MANN-WHITNEY

    (A-B) Localization of IFT-A proteins (A) IFT140 and (B) IFT43 in WT, WDR60 KO, and WDR34 KO#1 cells. Line graphs show lines scans of IFT intensity along the length of a representative cilium from WT (Ai and Bi, green), WDR60 KO (Aii and Bii, orange) and WDR34 KO (Aiii and Biii, yellow) cells. (C) Quantification of IFT140 intensity within cilia in WT and WDR60 KO cells (n=3, 186 WT and 166 WDR60 KO cells quantified). (D) Quantification of the number of IFT43 positive cilia from WT and WDR60 KO cells (n=3, 271 WT and 203 WDR60 KO cells quantified). (C-D) Mann-Whitney test was used, p-value: ****=<0.0001. (E) Localization of KAP3A in WT, WDR60 KO, and WDR34 KO cells. Scale bars = 5µm. Arrows point to the ciliary base.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A-B) Localization of IFT-A proteins (A) IFT140 and (B) IFT43 in WT, WDR60 KO, and WDR34 KO#1 cells. Line graphs show lines scans of IFT intensity along the length of a representative cilium from WT (Ai and Bi, green), WDR60 KO (Aii and Bii, orange) and WDR34 KO (Aiii and Biii, yellow) cells. (C) Quantification of IFT140 intensity within cilia in WT and WDR60 KO cells (n=3, 186 WT and 166 WDR60 KO cells quantified). (D) Quantification of the number of IFT43 positive cilia from WT and WDR60 KO cells (n=3, 271 WT and 203 WDR60 KO cells quantified). (C-D) Mann-Whitney test was used, p-value: ****=<0.0001. (E) Localization of KAP3A in WT, WDR60 KO, and WDR34 KO cells. Scale bars = 5µm. Arrows point to the ciliary base.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: MANN-WHITNEY

    (A and B) Single frame images are taken from live imaging movies of WT and WDR60 KO cells overexpressing EGFP-SSTR3 and GFP-Arl13b. (C-F) Fixed cell staining of overexpressed Arl13b-GFP, EGFP-SSTR3, EGFP-5HT6, and EGFP-Rab8a in WT and WDR60 KO cells. (Ci-Fi) Intensity quantification of the overexpressed protein indicated (Arl13b-GFP n=3, 56 WT and 50 WDR60 KO cells quantified; EGFP-SSTR3 n=3, 80 WT and 50 WDR60 KO cells quantified; EGFP-5HT6 n=3, 50 WT and 51 WDR60 KO cells quantified; EGFP-Rab8a n=3, 51 WT, and WDR60 50 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. Scale bars 5µ.m.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A and B) Single frame images are taken from live imaging movies of WT and WDR60 KO cells overexpressing EGFP-SSTR3 and GFP-Arl13b. (C-F) Fixed cell staining of overexpressed Arl13b-GFP, EGFP-SSTR3, EGFP-5HT6, and EGFP-Rab8a in WT and WDR60 KO cells. (Ci-Fi) Intensity quantification of the overexpressed protein indicated (Arl13b-GFP n=3, 56 WT and 50 WDR60 KO cells quantified; EGFP-SSTR3 n=3, 80 WT and 50 WDR60 KO cells quantified; EGFP-5HT6 n=3, 50 WT and 51 WDR60 KO cells quantified; EGFP-Rab8a n=3, 51 WT, and WDR60 50 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. Scale bars 5µ.m.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Imaging, Staining, MANN-WHITNEY

    (A-C) Localization of RPGRIP1L, TMEM67, and TCTN1 in WT and KO cells. (Ai and Bi) Percentage of RPGRIP1L and TMEM67 positive cilia. RPGRIP1L n=3, 188 WT and 272 WDR60 KO cells quantified; TMEM67 n=3, 359 WT and 243 WDR60 KO cells quantified. Mann-Whitney test was used p-value: ****=<0.0001. Scale bars 5µ.m. Arrows point to the ciliary base.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A-C) Localization of RPGRIP1L, TMEM67, and TCTN1 in WT and KO cells. (Ai and Bi) Percentage of RPGRIP1L and TMEM67 positive cilia. RPGRIP1L n=3, 188 WT and 272 WDR60 KO cells quantified; TMEM67 n=3, 359 WT and 243 WDR60 KO cells quantified. Mann-Whitney test was used p-value: ****=<0.0001. Scale bars 5µ.m. Arrows point to the ciliary base.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: MANN-WHITNEY

    (A and B) Immunofluorescence of WT and WDR60 KO cells in presence or absence of SAG and stained for Smo (green), AcTub (red), and DAPI (blue). (Ai and Bi) Percentage of Smo positive cilia in SAG untreated (n=3, 148 WT and 120 WDR60 KO cells quantified) and treated cells (n=3, 670 WT and 580 WDR60 KO cells quantified). (Bii) Quantification of the total intensity of ciliary Smo labeling in cells treated with SAG (n=3, 102 WT and 82 WDR60 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. Scale bars 5µ.m.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A and B) Immunofluorescence of WT and WDR60 KO cells in presence or absence of SAG and stained for Smo (green), AcTub (red), and DAPI (blue). (Ai and Bi) Percentage of Smo positive cilia in SAG untreated (n=3, 148 WT and 120 WDR60 KO cells quantified) and treated cells (n=3, 670 WT and 580 WDR60 KO cells quantified). (Bii) Quantification of the total intensity of ciliary Smo labeling in cells treated with SAG (n=3, 102 WT and 82 WDR60 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. Scale bars 5µ.m.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Immunofluorescence, Staining, Labeling, MANN-WHITNEY

    (A) Immunofluorescence staining of HA (green) in WDR60 KO cell lines stably expressing HA-tagged WT and mutant WDR60 constructs. (B and C) IFT88 or IFT140 staining (green) with AcTub labeling (red) of the stable cell lines shown in A as well as WT cells. (Bi) Total intensity quantification of IFT88 labeling across the length of primary cilia in each cell line (n=3; 97 WT, 125 WDR60 KO, 202 WDR60 KO+ HA-WDR60, 119 WDR60 KO+ HA-WDR60 [T749M], 150 WDR60 KO+ HA-WDR60 [Q631*] cells quantified). (Ci) Total intensity quantification of IFT140 labeling across the length of primary cilia in each cell line (n=3; 168 WT, 164 WDR60 KO, 63 WDR60 KO+ HA-WDR60, 56 WDR60 KO+ HA-WDR60 [T749M], 71 WDR60 KO+ HA-WDR60 [Q631*] cells quantified). (D and E) WDR34 KO#1 cells stably expressing mGFP-tagged WT and mutant WDR34. (D) Primary cilia staining with Arl13b (red) and AcTub (blue). (Di) Percentage of ciliated cells (n=3; 357 WT, 430 WDR34 KO, 399 WDR34 KO + mGFP-WDR34, 383 WDR34 KO + mGFP-WDR34 [A22V], 432 WDR34 KO + mGFP-WDR34[C148F] cells quantified). (E) IFT88 staining in WT, WDR34 KO#1 cells, and WDR34 KO#1 cells expressing GFP-tagged WT and mutant WDR34. One-way ANOVA followed by Kruskal-Wallis test was used p-value: ****=<0.0001. Arrows point to the ciliary base. (G-H) Immunoprecipitation of HA-tagged GFP, WT, WDR60 [T749M] and HA-WDR60 [Q631*] mutant proteins followed by immunoblot for (F) HA, NudCD3, and GAPDH; (G) WDR34, LIC3, and TCTEX1and (H) IFT140, IFT88, and IFT57.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A) Immunofluorescence staining of HA (green) in WDR60 KO cell lines stably expressing HA-tagged WT and mutant WDR60 constructs. (B and C) IFT88 or IFT140 staining (green) with AcTub labeling (red) of the stable cell lines shown in A as well as WT cells. (Bi) Total intensity quantification of IFT88 labeling across the length of primary cilia in each cell line (n=3; 97 WT, 125 WDR60 KO, 202 WDR60 KO+ HA-WDR60, 119 WDR60 KO+ HA-WDR60 [T749M], 150 WDR60 KO+ HA-WDR60 [Q631*] cells quantified). (Ci) Total intensity quantification of IFT140 labeling across the length of primary cilia in each cell line (n=3; 168 WT, 164 WDR60 KO, 63 WDR60 KO+ HA-WDR60, 56 WDR60 KO+ HA-WDR60 [T749M], 71 WDR60 KO+ HA-WDR60 [Q631*] cells quantified). (D and E) WDR34 KO#1 cells stably expressing mGFP-tagged WT and mutant WDR34. (D) Primary cilia staining with Arl13b (red) and AcTub (blue). (Di) Percentage of ciliated cells (n=3; 357 WT, 430 WDR34 KO, 399 WDR34 KO + mGFP-WDR34, 383 WDR34 KO + mGFP-WDR34 [A22V], 432 WDR34 KO + mGFP-WDR34[C148F] cells quantified). (E) IFT88 staining in WT, WDR34 KO#1 cells, and WDR34 KO#1 cells expressing GFP-tagged WT and mutant WDR34. One-way ANOVA followed by Kruskal-Wallis test was used p-value: ****=<0.0001. Arrows point to the ciliary base. (G-H) Immunoprecipitation of HA-tagged GFP, WT, WDR60 [T749M] and HA-WDR60 [Q631*] mutant proteins followed by immunoblot for (F) HA, NudCD3, and GAPDH; (G) WDR34, LIC3, and TCTEX1and (H) IFT140, IFT88, and IFT57.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Immunofluorescence, Staining, Stable Transfection, Expressing, Mutagenesis, Construct, Labeling, Immunoprecipitation, Western Blot

    (A) Immunoblotting for WDR60 and WDR34 in WT, WDR34 KO#1 and WDR60 KO cells. (B) LIC3 localization in the cilia of WT, WDR34 KO#1 and WDR60 KO cells. (C) DHC2 localization at the ciliary base in WT and KO cells. (Ci) Intensity quantification shows a reduction of DHC2 at the ciliary base in WDR34 KO#1 cells (n=3, 120 WT, 106 WDR60 KO and 71 WDR34 KO #1 cells quantified). (D) TCTEX1 localizes at the ciliary base in WT and KO cells (n=3 115 WT, 85 WDR60 KO, and 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5µm. Arrows point to the ciliary base.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: (A) Immunoblotting for WDR60 and WDR34 in WT, WDR34 KO#1 and WDR60 KO cells. (B) LIC3 localization in the cilia of WT, WDR34 KO#1 and WDR60 KO cells. (C) DHC2 localization at the ciliary base in WT and KO cells. (Ci) Intensity quantification shows a reduction of DHC2 at the ciliary base in WDR34 KO#1 cells (n=3, 120 WT, 106 WDR60 KO and 71 WDR34 KO #1 cells quantified). (D) TCTEX1 localizes at the ciliary base in WT and KO cells (n=3 115 WT, 85 WDR60 KO, and 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5µm. Arrows point to the ciliary base.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Western Blot, MANN-WHITNEY

    Abundance ratios in WT/ WDR34 KO cells expressing HA-WDR60 and WT/ WDR60 KO cells expressing HA-WDR34 determined by TMT analysis. Proteomic data were obtained from two independent experiments. The table shows raw data from one experiment. Similar results were obtained by normalizing the data with respect to the overexpressed protein abundance. (B) Schematic of the dynein-2 complex; the heavy chain is shown as two light blue homodimers, light intermediate chains are shown as hexagons, intermediate chains as extended ovals and light chains as circles. (Bi and Bii) Model of the dynein-2 complex in absence of (Bi) WDR60 or (Bii) WDR34. Reduced binding is represented by decreased color intensity. (Bi) In WDR60 KO cells interaction of HA-WDR34 with DHC2, DYNC2LI1/LIC3, DYNLT1/TCTEX1, DYNLL1/LC8-1, DYNLL2/LC8-2, TCTEX1D2, and DYNLRB1 is greatly reduced respect to WT cells. (Bii) In WDR34 KO cells interaction of HA-WDR60 with other components of dynein-2 is mostly unchanged.

    Journal: bioRxiv

    Article Title: Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

    doi: 10.1101/251694

    Figure Lengend Snippet: Abundance ratios in WT/ WDR34 KO cells expressing HA-WDR60 and WT/ WDR60 KO cells expressing HA-WDR34 determined by TMT analysis. Proteomic data were obtained from two independent experiments. The table shows raw data from one experiment. Similar results were obtained by normalizing the data with respect to the overexpressed protein abundance. (B) Schematic of the dynein-2 complex; the heavy chain is shown as two light blue homodimers, light intermediate chains are shown as hexagons, intermediate chains as extended ovals and light chains as circles. (Bi and Bii) Model of the dynein-2 complex in absence of (Bi) WDR60 or (Bii) WDR34. Reduced binding is represented by decreased color intensity. (Bi) In WDR60 KO cells interaction of HA-WDR34 with DHC2, DYNC2LI1/LIC3, DYNLT1/TCTEX1, DYNLL1/LC8-1, DYNLL2/LC8-2, TCTEX1D2, and DYNLRB1 is greatly reduced respect to WT cells. (Bii) In WDR34 KO cells interaction of HA-WDR60 with other components of dynein-2 is mostly unchanged.

    Article Snippet: The antibodies used, and their dilutions for western blotting (WB) and immunofluorescence (IF) are as follows: Acetylated tubulin (Sigma (Poole, UK) T6793 1:2000 for IF), rabbit anti-HA (Cell Signaling Technologies (New England Biolabs, Hitchin, UK) 1:2000 WB, 1:1000 IF), rabbit IFT88 (Proteintech (Manchester, UK) 13967-1-AP, 1:200 WB, 1:300 IF), rabbit anti-IFT140 (Proteintech 17460-1-AP, 1:200, WB 1:100 IF), rabbit anti-IFT57 (Proteintech 11083-1,-AP 1:200, WB 1:100 IF), rabbit anti-IFT43 (Proteintech 24338-1-AP, 1:50 IF), anti-IFT20 (Proteintech 13615-1-AP, 1:200 IF), anti-TMEM67 (proteintech13975-1-AP, 1:50 IF), anti-RPGRIP1L (Proteintech 55160-1-AP, 1:100 IF), anti-DYNC2HC1 (Proteintech 55473-1-AP, 1:100 IF), rabbit anti-LIC3 (Proteintech 15949-1-AP, 1:250 WB, 1:100 IF), rabbit anti-TCTEX1 (Santa Cruz Biotechnology (from Insight Biotechnology, Wembley, UK) sc-28537, 1:200 WB, 1:100 IF), rabbit anti-Arl13B (Proteintech 17711-1AP, 1:1000 IF), rabbit anti-TCTN1 (Proteintech 15004-1-AP, 1:100 IF), rabbit anti-Smo (Abcam (Cambridge, UK) ab38686, 1:100 IF), Sheep anti-Myc (( ) kindly provided by Harry Mellor, University of Bristol), rabbit anti-WDR60 (Novus Biologicals (from Bio-Techne Abingdon, UK) NBP1-90437 1:300 WB in ), rabbit anti-WDR60 (Sigma HPA021316, 1:300 WB in Fig. S2 and Fig.S5), rabbit anti-WDR34 (Novus NBP188805, 1:300 WB), mouse anti-GAPDH (Abcam ab9484, 1:1000 WB), p150 glued (BD 6127009, 1:1000 WB), LIS1 (Bethyl A300-409A, 1:1000 WB), dic74 (MAB1618 Millipore (Watford, UK), 1:1000 WB), NUDCD3 (Sigma HPA019136, 1:350 WB).

    Techniques: Expressing, Binding Assay

    Effect of baicalin on cellular apoptosis in mouse embryos. The nuclei of blastocysts (n = 30) were stained with Hoechst 33342 to examine cell apoptosis (Bar = 100 μm). Blastocysts from the in vitro (left), baicalin-treated (middle), and in vivo (right) groups showing higher degree of apoptosis of nuclei for in vitro group (white arrows) than the baicalin treated or in vivo group (A). Relative mRNA and protein expression levels of Caspase-3 (B), BAX (C) and BCL-2 (D), respectively, in mouse blastocysts (n = 30) from the in vitro , baicalin treated and in vivo groups. * P < 0.05 vs . in vivo control group and ** P < 0.01 vs . in vivo control group; # P < 0.05 vs . IVC group and ## P < 0.01 vs . IVC group.

    Journal: The Journal of Reproduction and Development

    Article Title: Baicalin increases developmental competence of mouse embryos in vitro by inhibiting cellular apoptosis and modulating HSP70 and DNMT expression

    doi: 10.1262/jrd.2016-047

    Figure Lengend Snippet: Effect of baicalin on cellular apoptosis in mouse embryos. The nuclei of blastocysts (n = 30) were stained with Hoechst 33342 to examine cell apoptosis (Bar = 100 μm). Blastocysts from the in vitro (left), baicalin-treated (middle), and in vivo (right) groups showing higher degree of apoptosis of nuclei for in vitro group (white arrows) than the baicalin treated or in vivo group (A). Relative mRNA and protein expression levels of Caspase-3 (B), BAX (C) and BCL-2 (D), respectively, in mouse blastocysts (n = 30) from the in vitro , baicalin treated and in vivo groups. * P < 0.05 vs . in vivo control group and ** P < 0.01 vs . in vivo control group; # P < 0.05 vs . IVC group and ## P < 0.01 vs . IVC group.

    Article Snippet: Goat polyclonal anti-HSP70 (sc-1060), rabbit polyclonal anti-BCL-2 (sc-492), and anti-BAX (sc-526) antibodies were obtained from Santa Cruz Biotechnology (USA), while rabbit polyclonal anti- Caspase-3 (ab-90437) antibodies were obtained from Abcam (Cambridge, UK).

    Techniques: Staining, In Vitro, In Vivo, Expressing