bioimagesUnknownMarta N. ShahbaziThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201306019JCB

Mouse keratinocytes were isolated from the backskin of p120 conditional knockout mice and grown in culture. After treatment with calcium for 6 h, cells were fixed and immunostained for ECadherin (red), CLASP2 (green), and p120 (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

CLASP2 localization was assessed in primary mouse keratinocytes treated with a scramble RNAi construct under low calcium conditions. Cells were immunostained for CLASP2 (red), α-tubulin (green), and DAPI (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were treated with calcium for 6 h and stained for CLASP2 (red), p120 (green), and DNA (DAPI, blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

p120-null (p120 Δ/Δ) mouse keratinocytes were transfected with p120ΔN-HA. Cells were treated with calcium for 6 h and immunostained for CLASP2 (red), p120 (green) and HA (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

p120-null (p120 Δ/Δ) mouse keratinocytes were treated with calcium for 6 h and immunostained for CLASP2 (red), p120 (green), ECadherin (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were stained for CLASP2 (red), ECadherin (green) and α-tubulin (blue) under normal calcium conditions. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Sections from the backskin of newborn wild-type mice were immunostained for or CLASP2 (red) together with p120 (green), DAPI (blue), and Integrin-β4 (white). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were treated with calcium for 24 h to allow the formation of differentiated suprabasal-like cells in vitro. Cells were immunostained for Nezha (red) together with ECadherin (green) and p120 (blue). In order to distinguish between basal and suprabasal cell-cell contacts, three different planes in the Z axis were imaged with a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were stained for CLASP2 (red), ECadherin (green) and α-tubulin (blue) under low calcium conditions the absence or presence of calcium. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Shown is a time-lapse microscopy video of the dynamic behavior of CLASP2 at cell-cell contacts. Wild-type primary mouse keratinocytes were transfected with plasmids encoding GFP-CLASP2 (green) and p120-cherry (red) and treated with calcium for 4-6 h. The dynamic behavior of both proteins at cell-cell contacts was analyzed by time-lapse microscopy. Images were taken at 37 ºC at intervals of 2 s in a TCS-SP5 (AOBS) laser scanning confocal microscope (Leica Microsystems) with a water immersion objective 63X HCX PL APO 1.2 N.A. using a temperature-controlled incubator chamber in the presence of CO2.

Wild-type (p120 f/f) keratinocytes were treated with calcium for 6 h and immunostained for CLASP2 (red), p120 (green), ECadherin (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Sections from the backskin of newborn wild-type mice were immunostained for Nezha (red) together with p120 (green), DAPI (blue), and Integrin-β4 (white). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were treated with calcium for 10 h following depolymerization of the microtubule network with a 30 µM nocodazole treatment for 2 h. Cells were immunostained for ECadherin (red), CLASP2 (green) and α-tubulin (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

CLASP2 expression was downregulated in primary mouse keratinocytes with lentiviruses expressing a specific shRNA. Cells were immunostained for CLASP2 (red), α-tubulin (green), and DAPI (blue) under low calcium conditions. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were treated with calcium for 10 h following depolymerization of the microtubule network with a 30 µM nocodazole treatment for 4 h. Cells were immunostained for ECadherin (red), CLASP2 (green) and α-tubulin (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were incubated with protein-A polystyrene beads coated with IgGs as a negative control for 5 h in the presence of calcium. Images show the localization of CLASP2 (red), p120 (green), DAPI (blue) and ECadherin (white). Brightfield images show the presence of the beads. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Mouse keratinocytes were isolated from the backskin of wild-type mice and grown in culture. After treatment with calcium for 6 h, cells were fixed and immunostained for ECadherin (red), CLASP2 (green), and p120 (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were treated with calcium for 24 h to allow the formation of differentiated suprabasal-like cells in vitro. Cells were immunostained for CLASP2 (red) together with ECadherin (green), and p120 (blue). In order to distinguish between basal and suprabasal cell-cell contacts, three different planes in the Z axis were imaged with a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Primary mouse keratinocytes were incubated with protein-A polystyrene beads coated with recombinant Fc-ECadherin for 5 h in the presence of calcium. Images show the localization of CLASP2 (red), p120 (green), DAPI (blue) and ECadherin (white). Brightfield images show the presence of the beads. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

CLASP2 localization was assessed in primary mouse keratinocytes treated with a scramble RNAi construct under normal calcium conditions. Cells were immunostained for CLASP2 (red), α-tubulin (green), and DAPI (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

Control primary mouse keratinocytes were treated with calcium for 10 h. Cells were immunostained for ECadherin (red), CLASP2 (green) and α-tubulin (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

CLASP2 expression was downregulated in primary mouse keratinocytes with lentiviruses expressing a specific shRNA. Cells were immunostained for CLASP2 (red), α-tubulin (green), and DAPI (blue) in the absence or presence of calcium. Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

p120-null (p120 Δ/Δ) mouse keratinocytes were transfected with p120FL-HA. Cells were treated with calcium for 6 h and immunostained for CLASP2 (red), p120 (green) and HA (blue). Images were taken on a Leica TCS-SP5 (AOBS) laser scanning confocal microscope.

bioimagesImage 575516 (Figure 8 - A)Figure 4 - CImage 575513 (Figure 5 - A)Image 575501 (Figure 1 - A)Image 575503 (Figure 1 - B)Image 575504 (Figure 1 - F)Figure 3 - AImage 575507 (Figure 3 - A)Video 1 - NoneImage 575522 (Figure 4 - C)Image 575508 (Figure 3 - E)Image 575502 (Figure 1 - A)Image 575514 (Figure 5 - A)Image 575518 (Figure 8 - D)Image 575517 (Figure 8 - A)Video 1Image 575509 (Figure 3 - E)Image 575524 (Video 1 - None)Figure 3Image 575506 (Figure 3 - A)Image 575510 (Figure 3 - E)Image 575519 (Figure 8 - D)Figure 1 - FImage 575520 (Figure 4 - C)Figure 1 - AFigure 5 - AImage 575512 (Figure 5 - A)Image 575515 (Figure 5 - A)Figure 1 - BFigure 3 - EImage 575511 (Figure 3 - E)Image 575505 (Figure 1 - F)Figure 8 - DFigure 8Figure 1Figure 5Figure 8 - AFigure 4Image 575521 (Figure 4 - C)Elaine FuchsMarta N. ShahbaziAnna AkhmanovaDiego MegiasGregg G. GundersenCarolina EpifanoMirna Perez-MorenofalseCLASP2 interacts with p120-catenin and governs microtubule dynamics at adherens junctions2013-12-23T11:25:27Z2018-11-29T11:25:27Z2018-11-29T11:25:27ZS-JCBD-20130601910.1083/jcb.201306019