Table 1. Key resources.

Reagent or resource	Source	Identifier
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.