Table 1.
Key resources
Reagent or resourceSourceIdentifier
Antibodies 
Rabbit anti-phospho Src (Tyr416; clone 2N8) EMD Millipore 04-857 
Rabbit anti-phospho FAK (Tyr397, clone 141-9) Invitrogen 44-625G 
Rabbit anti-phospho-paxillin (Tyr-118) ECM Biosciences PP4501 
Mouse anti-paxillin (clone 5H11) Invitrogen IAH00492 
Mouse anti-vinculin Sigma-Aldrich V9131 
Rabbit anti-APC Abcam ab1527 
Mouse anti-Dia1 (E-4) Santa Cruz Biotechnology sc-373807 
Rabbit anti-LC3 Novus Biologicals NB100-2220SS 
Donkey anti-rabbit Alexa Fluor 488 Thermo Fisher Scientific A-21206 
Goat anti-rabbit Alexa Fluor 555 Thermo Fisher Scientific A-21428 
Donkey anti-mouse Alexa Fluor 568 Thermo Fisher Scientific A-10037 
Rabbit anti-GFP Abcam ab6556 
Rabbit/human anti-GADPH Abcam ab9485 
Mouse/human anti-NBR1 Abnova H00004077-A01 
Donkey anti-rabbit IRDye 680RD Rockland Immunochemicals 926-68073 
Mouse anti-rabbit IRDye 680RD Rockland Immunochemicals 926-32220 
Bacterial and virus strains 
XL Blue competent cells This paper N/A 
Chemicals, peptides, and recombinant proteins 
Alexa Fluor 568–phalloidin Thermo Fisher Scientific A12380 
200 mM L-glutamine Thermo Fisher Scientific 25030-081 
DMEM Thermo Fisher Scientific 11995-073 
FBS Sigma-Aldrich F9423 
Lipofectamine 3000 Thermo Fisher Scientific L3000-015 
Lipofectamine RNAiMAX Thermo Fisher Scientific 13778075 
Hepes Sigma-Aldrich H4034 
100 mM sodium pyruvate Thermo Fisher Scientific 11360070 
Opti-MEM reduced serum media Thermo Fisher Scientific 31985-088 
Bafilomycin A Sigma Aldrich SML1661 
Pepstatin A US Biological P3280 
Collagen I Advanced BioMatrix CB-40236 
Formaldehyde 37% Sigma-Aldrich 252549 
Experimental models: Cell lines 
Human osteosarcoma cells: U2OS ATCC HTB96 
Human breast cancer cells: MDA-MB-231 ATCC HTB-26 
Oligonucleotides 
siRNA targeting sequence against the human APC: 5′-GGA​UCU​GUA​UCA​AGC​CGU​UTT-3′ sense and 5′-AAC​GGC​UUG​AUA​CAG​AUC​CTT-3′ antisense Juanes et al., 2017  N/A 
siRNA targeting sequence against the human Dia1: 5′-CUG​UUA​AUA​AAG​CAU​UGA​AUU-3′ sense and 5′-AAU​UCA​AUG​CUU​UAU​UAA​CAG-3′ antisense This paper N/A 
Stealth siRNA-control “scramble” oligos: 5′-CAG​UCG​CGU​UUG​CGA​CUG​G-3′ Invitrogen 12935-200 
Recombinant DNA 
Plasmid: APC-WT Juanes et al., 2017  Bruce Goode-US 
Plasmid: APC-m4 Juanes et al., 2017  Bruce Goode-US 
Plasmid: mCherry-zyxin Kenific et al., 2016  Addgene 55166 
Plasmid: pRK-GFP-paxillin Chen et al., 2013  Addgene 50529 
Plasmid: 3×GFP-EMTB Miller and Bement, 2009  Addgene 26742 
Plasmid: GFP-LC3 Jackson et al., 2005  Addgene 11546 
Plasmid: GFP-NBR1 Kenific et al., 2016  Addgene 74202 
Plasmid: mCherry-LAMP1 Miyamoto et al., 2015  Addgene 61524 
Plasmid: GFP-LAMP1 Falcón-Pérez et al., 2005  Addgene 34831 
Plasmid: GFP-actin Svitkina et al., 2003  Tatyana Svitkina-US 
Plasmid: GFP empty vector This paper Addgene 54522 
Premo Autophagy Tandem Sensor RFP-GFP-LC3B Thermo Fisher Scientific P36239 
Software and algorithms 
Fiji/ImageJ Schindelin et al., 2012  N/A 
NIS Elements software, version 4.30.02 Nikon Instruments N/A 
Metamorph Software 7.7.8 Molecular Devices N/A 
MATLAB 2017 MathWorks N/A 
FAAS Berginski et al., 2011; Berginski and Gomez, 2013  N/A 
GraphPad Prism 6.0c GraphPad Software N/A 
Illustrator CS6 Adobe Systems N/A 
Filament sensor Eltzner et al., 2015  N/A 
MATLAB (polarization-resolved fluorescence analysis algorithm) MathWorks (Kress et al., 2013) and this paper N/A 
Labview (acquisition software for polarization-resolved fluorescence imaging) National Instruments (Wang et al., 2013) N/A 
Odyssey Infrared Imaging System LI-COR Biotechnology N/A 
Other 
AquaMount mounting media Thermo Fisher Scientific 14-390-5 
Prolong Diamond Antifade Mounting Thermo Fisher Scientific P-36961 
GFP–Trap-A agarose beads ChromoTek gta-20 
Micro cover glasses 22 × 22 mm VWR International PA 48366-067 
Circular Round cover glasses 0.15 mm Thermo Fisher Scientific 12-545-80 
High precision cover glasses 1.5H 22 × 22 mm Azer Scientific ES0107052 
Dishes with 35-mm glass bottom Thermo Fisher Scientific P-35-1.5-14-C 
Glass pattern coverslips (starter’s CYTOO) CYTOO 10-900-00 
Reagent or resourceSourceIdentifier
Antibodies 
Rabbit anti-phospho Src (Tyr416; clone 2N8) EMD Millipore 04-857 
Rabbit anti-phospho FAK (Tyr397, clone 141-9) Invitrogen 44-625G 
Rabbit anti-phospho-paxillin (Tyr-118) ECM Biosciences PP4501 
Mouse anti-paxillin (clone 5H11) Invitrogen IAH00492 
Mouse anti-vinculin Sigma-Aldrich V9131 
Rabbit anti-APC Abcam ab1527 
Mouse anti-Dia1 (E-4) Santa Cruz Biotechnology sc-373807 
Rabbit anti-LC3 Novus Biologicals NB100-2220SS 
Donkey anti-rabbit Alexa Fluor 488 Thermo Fisher Scientific A-21206 
Goat anti-rabbit Alexa Fluor 555 Thermo Fisher Scientific A-21428 
Donkey anti-mouse Alexa Fluor 568 Thermo Fisher Scientific A-10037 
Rabbit anti-GFP Abcam ab6556 
Rabbit/human anti-GADPH Abcam ab9485 
Mouse/human anti-NBR1 Abnova H00004077-A01 
Donkey anti-rabbit IRDye 680RD Rockland Immunochemicals 926-68073 
Mouse anti-rabbit IRDye 680RD Rockland Immunochemicals 926-32220 
Bacterial and virus strains 
XL Blue competent cells This paper N/A 
Chemicals, peptides, and recombinant proteins 
Alexa Fluor 568–phalloidin Thermo Fisher Scientific A12380 
200 mM L-glutamine Thermo Fisher Scientific 25030-081 
DMEM Thermo Fisher Scientific 11995-073 
FBS Sigma-Aldrich F9423 
Lipofectamine 3000 Thermo Fisher Scientific L3000-015 
Lipofectamine RNAiMAX Thermo Fisher Scientific 13778075 
Hepes Sigma-Aldrich H4034 
100 mM sodium pyruvate Thermo Fisher Scientific 11360070 
Opti-MEM reduced serum media Thermo Fisher Scientific 31985-088 
Bafilomycin A Sigma Aldrich SML1661 
Pepstatin A US Biological P3280 
Collagen I Advanced BioMatrix CB-40236 
Formaldehyde 37% Sigma-Aldrich 252549 
Experimental models: Cell lines 
Human osteosarcoma cells: U2OS ATCC HTB96 
Human breast cancer cells: MDA-MB-231 ATCC HTB-26 
Oligonucleotides 
siRNA targeting sequence against the human APC: 5′-GGA​UCU​GUA​UCA​AGC​CGU​UTT-3′ sense and 5′-AAC​GGC​UUG​AUA​CAG​AUC​CTT-3′ antisense Juanes et al., 2017  N/A 
siRNA targeting sequence against the human Dia1: 5′-CUG​UUA​AUA​AAG​CAU​UGA​AUU-3′ sense and 5′-AAU​UCA​AUG​CUU​UAU​UAA​CAG-3′ antisense This paper N/A 
Stealth siRNA-control “scramble” oligos: 5′-CAG​UCG​CGU​UUG​CGA​CUG​G-3′ Invitrogen 12935-200 
Recombinant DNA 
Plasmid: APC-WT Juanes et al., 2017  Bruce Goode-US 
Plasmid: APC-m4 Juanes et al., 2017  Bruce Goode-US 
Plasmid: mCherry-zyxin Kenific et al., 2016  Addgene 55166 
Plasmid: pRK-GFP-paxillin Chen et al., 2013  Addgene 50529 
Plasmid: 3×GFP-EMTB Miller and Bement, 2009  Addgene 26742 
Plasmid: GFP-LC3 Jackson et al., 2005  Addgene 11546 
Plasmid: GFP-NBR1 Kenific et al., 2016  Addgene 74202 
Plasmid: mCherry-LAMP1 Miyamoto et al., 2015  Addgene 61524 
Plasmid: GFP-LAMP1 Falcón-Pérez et al., 2005  Addgene 34831 
Plasmid: GFP-actin Svitkina et al., 2003  Tatyana Svitkina-US 
Plasmid: GFP empty vector This paper Addgene 54522 
Premo Autophagy Tandem Sensor RFP-GFP-LC3B Thermo Fisher Scientific P36239 
Software and algorithms 
Fiji/ImageJ Schindelin et al., 2012  N/A 
NIS Elements software, version 4.30.02 Nikon Instruments N/A 
Metamorph Software 7.7.8 Molecular Devices N/A 
MATLAB 2017 MathWorks N/A 
FAAS Berginski et al., 2011; Berginski and Gomez, 2013  N/A 
GraphPad Prism 6.0c GraphPad Software N/A 
Illustrator CS6 Adobe Systems N/A 
Filament sensor Eltzner et al., 2015  N/A 
MATLAB (polarization-resolved fluorescence analysis algorithm) MathWorks (Kress et al., 2013) and this paper N/A 
Labview (acquisition software for polarization-resolved fluorescence imaging) National Instruments (Wang et al., 2013) N/A 
Odyssey Infrared Imaging System LI-COR Biotechnology N/A 
Other 
AquaMount mounting media Thermo Fisher Scientific 14-390-5 
Prolong Diamond Antifade Mounting Thermo Fisher Scientific P-36961 
GFP–Trap-A agarose beads ChromoTek gta-20 
Micro cover glasses 22 × 22 mm VWR International PA 48366-067 
Circular Round cover glasses 0.15 mm Thermo Fisher Scientific 12-545-80 
High precision cover glasses 1.5H 22 × 22 mm Azer Scientific ES0107052 
Dishes with 35-mm glass bottom Thermo Fisher Scientific P-35-1.5-14-C 
Glass pattern coverslips (starter’s CYTOO) CYTOO 10-900-00 

N/A, not applicable.

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