Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a , b ′ E-Cadherin-labelled placodes of control and γ-tubulin-overexpressing embryos. Magenta boxed areas in a , b are magnified in a ′ , b ′. Dotted lines denote placode boundary, asterisks the invagination pit. Magenta arrows in b ′ point to wavy junctions in ncd:: γ -tubulin-EGFP embryos. c–e Quantification of junction waviness. c Placodes of ncd:: γ -tubulin-EGFP embryos show a significantly higher proportion of wavy junctions than control placodes (7 placodes for control and 10 placodes for ncd:: γ -tubulin-EGFP were analysed); p = 0.0001, determined by two-sided unpaired Mann–Whitney t -test, shown are mean ± SD. d Wavy junctions in ncd:: γ -tubulin-EGFP overexpressing embryos are significantly less straight (90 junctions from 5 placodes) than randomly picked junctions in control placodes (90 junctions from 5 placodes). Statistical significance was determined using two-sided unpaired Mann–Whitney t -test with p < 0.0001, shown are violin plots with median and quartiles. e The straightness of a junction is defined as the ratio of the length of the junction itself (L [junction] ) divided by the length of the direct route between vertices (L [direct route] ) , . For a straight junction this value is close to 1, for a wavy junction it is «1. f Apices of secretory cells of ncd:: γ -tubulin-EGFP embryos are more constricted than apices of control placodes, illustrated are both percentage of cells of a certain apical area bin as well as the cumulative percentage of cells. 738 cells were analysed in 6 ncd:: γ -tubulin-EGFP embryos and 425 cells in 4 control placodes; Kolmogorov–Smirnov-two-sample test on the cumulative data did not show a significant difference between control and γ-tubulin overexpressing embryos ( p = 0.6068). However, comparing the distribution of cells with small apical areas between 0 and 5 μm 2 ( p = 0.0361) and cells with apical areas in a range between 5 and 10 μm 2 ( p = 0.0361), showed a significant difference, indicated by green shaded area. g ncd:: γ -tubulin-EGFP embryos show increased apical-medial myosin compared to control (visualised using sqh-RFP ). 100 cells in 4 placodes were analysed in ncd:: γ -tubulin-EGFP embryos and 125 cells in five control placodes; statistical significance was deduced by two-sided unpaired Mann-Whitney test as p < 0.0001, shown is mean ± SD. h , i Microtubules within the apical-medial region of placodal cells of ncd:: γ -tubulin-EGFP embryos remain organised in a non-centrosomal fashion ( i ), as in the control ( h ). Arrowheads point to ends of microtubules or microtubule bundles away from centrosomes. j–l An increased amount of apica-medial Patronin-RFP, a stabiliser of free microtubule minus-ends, accumulates in placodal cells of ncd:: γ -tubulin-EGFP embryos where increased numbers of microtubule bundle foci are found ( k ) in comparison to control ( j ), indicated by blue arrowheads. l Quantification of medial to junctional Patronin-RFP intensity; 298 cells in 10 placodes were analysed in ncd:: γ -tubulin-EGFP embryos and 349 cells in 11 control placodes; statistical significance was deduced by two-sided unpaired Mann–Whitney test as p < 0.0001, shown is mean ± SD. m Model of the effect of γ-tubulin overexpression in placodal cells. See also Supplementary Fig. .

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Control, MANN-WHITNEY, Comparison, Over Expression

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a Schematic of the ‘degradFP’ tissue-specific-degradation system (as in ref. ): tissue-specific expression of an F-box/anti-GFP-nanobody fusion protein, degradFP, (using UAS/Gal4) leads to tissue-specific degradation of any endogenously GFP/YFP-tagged protein, in this case Katanin80-YFP. b – d Expressing degradFP using fkh-Gal4 in the salivary gland placode ( Kat80YFP degradFP fkhG4 ; c – c″ ) leads to significant loss of Katanin80-YFP compared to control ( Kat80YFP degradFP ctrl ; b – b″ ). Cell outlines are marked by E-Cadherin (E-Cad). d Quantification of Katanin80-YFP depletion ( Kat80YFP degradFP ctrl n = 33 embryos; Kat80YFP degradFP fkhG4 n = 34 embryos; shown are mean ± SD, statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001). b″ and c″ are higher magnifications of the white boxes marked in b and c , respectively. e – g In placodes where Katanin80-YFP is degraded ( f , f ′), microtubules (green, labelled for acetylated α-tubulin) remain localised within the apical domain and in contact with centrosomes (magenta, labeled for Asl) compared to control ( e , e ′) where a non-centrosomal longitudinal array is formed. e ′ and f ′ are magnifications of the areas indicated in e and f by a white box. g Quantification of the effect shown in e , f ; ( Kat80YFP degradFP ctrl : 440 cells from 14 embryos; Kat80YFP degradFP fkhG4 : 541 cells from 20 embryos; shown are mean ± SD, statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001). h - h‴ Katanin80-YFP degradation ( h ′) leads to a loss of apical constriction compared to control ( h ), apical area of cells of example placodes are shown in a heat map. h″ Quantification of apical area distribution of placodal cells in control (ctrl) and Katanin depleted ( degradFP fkhGal4 ) placodes at stage 11 showing the cumulative percentage of cells relative to apical area size. h‴ Percentage of cells in different size-bins [Kolmogorov-Smirnov two-sample test, p « 0.001 (***)]. 12 placodes were segmented and analysed for control and 13 for Katanin80-YFP depletion, the total number of cells traced was N( Kat80YFP degradFP ctrl )=1373, N( Kat80YFP degradFP fkhG4 ) = 1162. i – k In placodes where Katanin80-YFP is degraded ( j , j ′), apical-medial F-actin (green, labeled using phalloidin) is reduced compared to control ( i , i ′) where apical-medial actin is highly prevalent. i ′ and j ′ are magnifications of the areas indicated in i and j by a white box. k Quantification of loss of apical-medial F-actin ( Kat80YFP degradFP ctrl: 924 cells from 17 embryos; Kat80YFP degradFP fkhG4: 919 cells from 15 embryos); shown are mean ± SD, statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001. The salivary gland placode is indicated by a white dotted line and the invagination point, where present, by an asterisk.

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Expressing, Control, MANN-WHITNEY, Labeling

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a–c ′ Patronin-YFP (green and a′ , b , c ), accumulates at adherens junctions (magenta) throughout the embryonic epidermis ( a and c , c ′), but in the apically constricting cells of the placode accumulates in an apical-medial position ( a and b , b ′). b and c ′ are magnification of the white boxes in a . d , e Apical-medial Patronin-YFP localises to the minus-ends of longitudinal microtubules labeled for tyrosinated α-tubulin. e is a magnification of the white box in d and also shows corresponding xz and yz-cross-sections. f – h Patronin depends on microtubules for its apical-medial localisation. In contrast to control ( f – f ″) where Patronin-YFP localises to apical-medial sites, when microtubules are lost upon expression of UAS-Spastin under fkh-Gal4 control ( g – g″ ), Patronin-YFP continues to be localised to junctional sites and does not relocalise to apical-medial sites. h Quantification of reduction of apical-medial Patronin-YFP upon placodal microtubule loss ( fkhGal4 ctrl: 955 cells from 13 embryos; fkhGal4 x UAS-Spastin: 999 cells from 19 embryos/ shown are mean ± SD, statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001). i – m Stills of time lapse movies of laser-induced microtubule ablation. Compared to a control cut just below the apical microtubules ( i ), laser-induced ablation of microtubules within the apical microtubule array in early stage 11 placodes ( j ) as well as in the apical region of the longitudinal microtubule array in later stage 12 placodes ( k ) leads to a very rapid recruitment of Patronin-RFP to the newly generated minus-ends of microtubules. Orange arrowheads in j , k point to the severed site and Patronin-RFP recruitment, blue arrows in i point to the control cut position. A Jupiter-GFP Patronin-RFP genotype was analysed in all instances. The dotted boxes indicate the area shown in individual black and white panels. See also Supplementary Fig. and Supplementary Movies – . l Time-resolved quantification of the fluorescence intensity of Patronin-RFP at the site of laser-ablation in early and late placodes compared to control. Mean ± SEM are shown, n = 17 (early placodes), n = 22 (late placodes), n = 22 (control). m Quantification of Patronin-RFP intensity in the first image acquired post-ablation in early and late placodes and control. Shown are mean ± SD; n = 17 (early placodes), n = 22 (late placodes), n = 22 (control); statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001 where indicated. The salivary gland placode is indicated by dotted lines and the invagination point by asterisks.

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Labeling, Control, Expressing, MANN-WHITNEY, Generated, Fluorescence

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a–d ″ Patronin-YFP ( a ′, b ″, c ′, d ″ and green in a , c ) and acetylated α-tubulin ( a ″, c ″ and magenta in a , b , b ′, c , d , d ′) labelling in control embryos ( a , b ″) and UAS-Patronin-RNAi x DaGal4 embryos ( c , d ″). E-Cadherin to label cell outlines is in yellow in a , b , green in b , b ′, d , d ′. b ′ and d ′ are z-sections of cells at the positions of the dotted lines indicated in b and d . See quantification in Supplementary Fig. . e – g Depletion of Patronin using RNAi in UAS-RNAi-Patronin x DaGal4 embryos ( f , f ′) leads to loss of apical-medial F-actin labelled using phalloidin (magenta) in contrast to control ( e , e ′). e ′ and f ′ are magnifications of the white boxes shown in e and f . Membranes are labelled by E-Cadherin (green). g Quantification of changes of apical-medial F-actin accumulation upon Patronin depletion; control: 316 cells from 9 embryos; UAS-RNAi-Patronin x DaGal4: 253 cells from 7 embryos; shown are mean ± SD, statistical significance was determined by two-sided unpaired Mann–Whitney test as p < 0.0001. h – j ′ Depletion of Patronin in UAS-Patronin-RNAi x DaGal4 embryos ( i ) leads to a loss of apical constriction compared to control ( h ), apical area of cells of example placodes are shown in a heat map. j , j ′ Quantification of apical area distribution of placodal cells in control and Patronin-depleted ( UAS-Patronin-RNAi x DaGal4 ) placodes at stage 11 showing the cumulative percentage of cells relative to apical area size ( j ) as well as the percentage of cells in different size-bins ( j ′) [Kolmogorov–Smirnov two-sample test, p « 0.001 (***)]. Six placodes were segmented and analysed for each condition, the total number of cells traced was N ( control ) = 585, N ( UAS-Patronin-RNAi x DaGal4 )=399. k Model of generation of the longitudinal non-centrosomal microtubule array in salivary gland placodal cell prior to apical constriction: inactivation of the Centrobin-depleted centrosome concomitant with entering cycle 17 and becoming post-mitotic leads to restriction of microtubule nucleation to a single centrosome. An increase in apical-medial Katanin levels in the secretory cells drive severing of microtubules at the active centrosome. Severed microtubule minus-ends are then captured by apical Patronin in the apical-medial region, thereby promoting the longitudinal microtubule arrangement. In overview panels the salivary gland placode is indicated by a dotted line and the invagination point by an asterisk.

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Control, MANN-WHITNEY

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a Centrosomes are usually built of two centrioles, a mother centriole (M) inherited from the last division, and a newly nucleated daughter centriole (D), surrounded by a cloud of PCM. Key centriole components: Asl, Spd-2, Sas-4, Centrobin (only on daughter centriole); PCM components: Polo-kinase, Cnn and γ-tubulin. b , c′ Whereas the centriole components Asl, Spd-2 and Sas-4 are equally enriched on both centrosomes in placodal cells, PCM components Polo, Cnn and γ-tubulin show asymmetric accumulation at placodal centrosomes (localisation for all in b and quantification in c , c′ ). Cell outlines are marked by E-Cadherin (E-Cad) or aPKC in b . c , c′ show the same data, with c′ showing a zoomed version of the 0–20 values of the asymmetry index of c . n values are: Sas4-GFP : 170 cells from 5 embryos; Spd-2-GFP : 139 cells from 4 embryos; Asl (antibody, ab): 302 cells from 9 embryos; GFP-Polo : 270 cells from 7 embryos; γ-tubulin (ab): 487 cells from 12 embryos; Cnn (ab): 309 cells from 9 embryos; Cnb-YFP : 157 cells from 5 embryos. Pairwise comparison of distributions was done via Kruskal–Wallis test (one-way ANOVA, all p values of these are listed in the “Methods” section). Mean and 95% CI are shown. Here and in all following quantifications cell ( c ) and embryo ( e ) numbers are indicated below the plots. d , e 90% of Cnn-containing centrosomes actively nucleate microtubules. d Projection of 30 consecutive time frames, 0.55 s apart, of a time lapse movie of EB1-GFP Cnn-RFP flies. d′ shows the Cnn-RFP channel to indicate the positions of centrosomes. d ″ schematically illustrates the EB1 comets moving away from Cnn-positive centrosomes that indicate active microtubule nucleation. e Quantification of nucleation capacity of 70 Cnn-positive centrosomes. See Supplementary Movie . f – g Cnn and Centrobin (Cnb) accumulate asymmetrically in individual cells, but on the same centrosome. Box in f is shown enlarged in f′ – f″″ . Asl labels all centrosomes and E-Cad labels cell outlines in f′ – f″″ . g Line scan profile through both centrosomes of a single cell illustrates the co-enrichment of Cnb and Cnn on the same centrosomes. More line scan examples are shown in Supplementary Fig. . The salivary gland placode is indicated by a dotted line and the invagination point by an asterisk in f . Filled arrowheads in b and f′ – f″″ indicate centrosomes with PCM and Centrobin accumulation, respectively, while hollow arrowheads indicate centrosomes without PCM or Centrobin staining. See also Supplementary Fig. .

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Comparison, Staining

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a , b Overexpression of γ-tubulin using ncd:: γ -tubulin-EGFP leads to reduction of γ-tubulin asymmetry at centrosomes (localisation in a– a″ and quantification in b ). b ncd:: γ -tubulin-EGFP asymmetry was quantified as part of the data illustrated in Fig. ′. The γ-tubulin (ab) data are reproduced here in comparison to the ncd:: γ -tubulin-EGFP overexpression: with n = 284 cells from 8 embryos; statistical significance was deduced by two-sided Mann–Whitney test of comparison as p < 0.0001, shown are mean and 95% CI. c Overexpression of ncd:: γ -tubulin-EGFP and loss of γ-tubulin asymmetry leads to increased levels of microtubule labelling using antibodies against α-tubulin or tyrosinated α-tubulin, but a slight reduction in acetylated α-tubulin labelling. Analysed were: 12 embryos for control and 19 embryos for ncd:: γ -tubulin-EGFP for α-tubulin, 18 embryos for control and 25 embryos for ncd:: γ -tubulin-EGFP fortyrosinated α-tubulin; 14 embryos for control and 18 embryos for ncd:: γ -tubulin-EGFP for acetylated α-tubulin. Statistical significance was determined by two-sided unpaired t -tests with Welch’s correction for α-tubulin ( p < 0.0001) and tyrosinated α-tubulin ( p < 0.0001) and a two-sided Mann–Whitney test for acetylated α-tubulin ( p = 0.007), shown are mean ± SD. d – f′ Labelling of stage 11 placodes of control and γ-tubulin overexpressing embryos, showing the increase in tyrosinated α-tubulin ( d , d′ ) and phalloidin labelling to reveal F-actin ( f , f′ ) and the decrease in acetylated α-tubulin labelling ( e , e′ ). See also Supplementary Fig. .

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Over Expression, Comparison, MANN-WHITNEY, Control

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet: a–a″ Katanin80, labelled using a YFP-protein trap line (green in a , a″ and single channel in a ′), accumulates specifically in the secretory placodal cells and not the surrounding epidermis. a″ Katanin80-YFP foci are found in an apical-medial position in the constricting population of cells (green arrows). Cell outlines are marked by E-Cadherin (magenta in a , a″ ). The salivary gland placode is indicated by a white dotted line and the invagination point by an asterisk. b , c Katanin80-YFP accumulates near centrosomes and microtubules. b Kat80YFP in green localises close to centrosomes marked by Asl-mCherry in red, cell outlines in white are labelled by E-Cadherin. b ′ Quantification of Katanin80-YFP accumulation near centrosomes. c Katanin80-YFP (green) can be found near microtubules labelled by staining for acetylated α-tubulin (white) in the proximity of centrosomes marked by Asl-mCherry (red). d – d ′ Katanin80-YFP (green) accumulates near the microtubule-nucleating centrosomes marked by Cnn (yellow), with all centrosomes marked by Asl-mCherry (cyan) and cell outlines marked by Crumbs (magenta). Solid arrowheads indicate centrosomes exhibiting stronger Cnn and Katanin accumulation, while hollow arrowheads point towards centrosomes with less or no Cnn and Katanin staining. d ′ Line scan profile through both centrosomes of a single cell to illustrate the co-enrichment of Katanin80-YFP on the Cnn-enriched centrosome. Centrosome positions are marked by Asl-mCherry. e , f Live imaging of microtubules (labeled with Jupiter-GFP, a microtubule-binding protein, in green) and centrosomes marked by Asl-mCherry (magenta) reveals microtubule release from centrosomes, two examples are shown. Arrowheads point to Jupiter foci that lose association with the centrosome over the course of the movie.

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Staining, Imaging, Labeling, Binding Assay

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet:

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: Control

Journal: Nature Communications

Article Title: A release-and-capture mechanism generates an essential non-centrosomal microtubule array during tube budding

doi: 10.1038/s41467-021-24332-0

Figure Lengend Snippet:

Article Snippet: Anti-E-Cadherin (DCAD2, 1:10 dilution), anti-CrebA (CrebA Rbt-PC, 1:1000) and anti-Crumbs (Cq4, 1:10) antibodies were obtained from the Developmental Studies Hybridoma Bank at the University of Iowa; anti-aPKC (sc-216; Santa Cruz) anti tyrosinated α-tubulin (YL1/2, 1:10) ; anti-acetylated α-tubulin (clone 6-11B-1, 1:500; Sigma); anti-α-tubulin (DM1A, 1:1000; Sigma); anti-γ-tubulin (clone GTU-88, 1:500; Sigma); anti-phospho-Histone H3 [Ser10] (Cell Signalling Technology; #9701, 1:500); anti-Asterless-NT; and anti-Cnn (1:1000n for both) were a kind gift from Jordan Raff ; anti-Shot (1:1000).

Techniques: CRISPR