bioimagesUnknownMark T. HowesThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201002119JCB

20 mgmL HRP uptake in Cav1-/- MEFs for 15 seconds in the presence of unlablled CTxB.

Fractions 2.8 was fixed onto EM grids and labeled for Cav1.

CTxB-555 and Tf-647 were pulsed into untreated Cav1-/- MEFs for 5 minutes, cells fixed and labeled with CTxB-488 post fixation.

CTxB-HRP was pulsed into Cav1-/-MEFs for 15 seconds, DAB reaction performed and cells HPF, freeze substituted and processed for EM.

WT MEFs were wounded and cells allowed to migrate into wound for 4-8 hours. Cells were pulsed for 2 minutes with CTxB, Tf and anti-CD44 antibodies.

Anti-CD44-HRP was pulsed into Cav1-/- MEFs for 2 minutes, DAB reaction performed and cells fixed and processed for EM.

CTxB-HRP was pulsed into Dyngo4a treated Cav1-/- MEFs for 2 minutes, DAB reaction performed and samples processed for EM.

WT MEFs were wounded and allowed to migrate for 4 hours. CTxB-HRP was pulsed into cells for 2 minutes, DAB reaction performed and cells fixed and processed for EM.

Anti-Thy-1, CTxB-555 and Tf-647 were pulsed into Cav1-/- MEFs for 2 minutes prior to fixation.

CTxB-HRP was added to cells for 5 mintues prior to fractionation. Fraction 2.8 was fixed and adsorbed onto EM grids and DAB reaction performed.

CTxB-555 and Tf-647 were pulsed into Cav1-/- MEFs for 2 minutes prior to fixation and labeling for myoferlin

Fractions 2.8 was fixed onto EM grids and labeled for Flot1.

20 mgmL HRP uptake in Cav1-/- MEFs without any CTxB.

Migrating WT MEFs were treated with Nocodazole for 30 minutes prior to addition of CTxB-555, Tf-647 and anti-CD44 antibodies for 2 minutes.

CTxB-HRP was pulsed into untreated Cav1-/- MEFs for 2 minutes, DAB reaction performed and samples processed for EM.

Anti-CD44-HRP was pulsed into Cav1-/- MEFs for 10 minutes, DAB reaction performed and cells fixed and processed for EM.

WT MEFs were wounded and cells allowed to migrate into the wound for 4-8 hours. Cells were pulsed for 2 minutes with CTxB-555 and Tf-647 and were labeled post fixation for Cav1.

Fractions 2.8 was fixed onto EM grids and labeled for Dynamin.

GFP-GPI cells were treated with wortmannin for 20 minutes prior to addition of anti-GFP antibodies for a further 10 minutes.

Anti-CD44-HRP was pulsed into Cav1-/- MEFs for 40 minutes, DAB reaction performed and cells fixed and processed for EM.

Fractions 2.8 was fixed onto EM grids and labeled for GRAF1.

Migrating WT MEFs were treated with Taxol for 30 minutes prior to addition of CTxB-555, Tf-647 and anti-CD44 antibodies for 2 minutes.

EM of membranes in Fraction 2.8

Anti-CD44, CTxB-555 and Tf-647 were pulsed into Cav1-/- MEFs for 2 minutes prior to fixation.

CTxB-555 and Tf-647 were pulsed into Dyngo4a Cav1-/- MEFs for 5 minutes, cells fixed and labeled with CTxB-488 post fixation.

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B)Image 13314 (Figure 5 - E)Image 13245 (Figure 4 - B)Image 13342 (Figure 1 - G)Image 13362 (Figure 4 - D)Image 13338 (Figure 1 - G)Image 13276 (Figure 4 - G)Image 13233 (Figure 3 - B)Image 13403 (Figure 5 - A)Image 13374 (Figure 6 - C)Image 13194 (Figure 1 - A)Image 13229 (Figure 3 - B)Image 13385 (Figure 6 - C)Image 13349 (Figure 1 - G)Image 13399 (Figure 5 - B)Image 13321 (Figure 6 - A)Image 13280 (Figure 4 - G)Figure 6 - CFigure 6 - AImage 13317 (Figure 6 - A)Image 13353 (Figure 1 - G)Figure 6 - EJohn F. HancockPiers J. WalserRumasia BashirRobert G. PartonAlun JonesAdam McCluskeySatyajit MayorRichard LundmarkMargaret R. LindsayHarvey McMahonKatia CortesePhillip J. RobinsonGordana HadzicMatthew KirkhamMichelle M. HillFiona SimpsonLibin LiuMark T. HowesDelia J. Hernandez-DeviezJames RichesPaul PilchfalseClathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells2010-08-16T11:19:54Z2018-11-29T11:19:54Z2018-11-29T11:19:54ZS-JCBD-20100211910.1083/jcb.201002119