Protocol detail

Autophagy and Actin Dynamics Regulation

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pN3-3xFlag-Control (Addgene, 107717, Guntram Suske Lab [75 (link)]); pmRFP-LC3 (Addgene, 21075, Tamotsu Yoshimori Lab [43 (link)]); pMXs-puro GFP-Sqstm1/p62 (Addgene, 38277, Noboru Mizushima Lab [49 (link)]); pBABE-puro mCherry-EGFP-LC3B (Addgene, 22418, Jayanta Debnath Lab [76 (link)]); pBABE-EGFP (Addgene, 36999, Debu Chakravarti Lab [77 (link)]); ECFP-SSH1ΔC (N461) (Dr. Mizuno lab [51 (link)]); EGFP-SSH1 (Dr. Storz lab [69 (link)]); mKeima-Red-Mito-7 (Addgene, 56018); pOPTN-EGFP (Addgene, 27052Beatrice Yue Lab [78 (link)]); pMXs-puro GFP-Sqstm1/p62ΔC (Addgene, 38282, Noboru Mizushima Lab [49 (link)]); pMXs-puro GFP-Sqstm1/p62 D337, 338, 339A (GFP-SQSTM1-LIR) (Addgene, 38280, Noboru Mizushima Lab [49 (link)]); HA-Ubiquitin (Addgene, 18712, Edward Yeh Lab [79 (link)]); pDR125 (Addgene, 37150, Dale Ramsden Lab); pN3-3xFlag-SSH1, mCherry-EGFP-Sqstm1/p62, mCherry-EGFP-Sqstm1^S403A, mCherry-EGFP-Sqstm1^S403E, pN3-Flag-SSH1ΔC, pN3-Flag-SSH1ΔN and pN3-Flag-SSH1ΔN^C393S were generated in this work (see methods below).

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Fang C., Woo J.A., Liu T., Zhao X., Cazzaro S., Yan Y., Matlack J., Kee T., LePochat P, & Kang D.E. (2020). SSH1 impedes SQSTM1/p62 flux and MAPT/Tau clearance independent of CFL (cofilin) activation. Autophagy, 17(9), 2144-2165.

Publication 2020

pN3-3xFlag-Control pmRFP-LC3 pBABE-puro mCherry-EGFP-LC3B pBABE-EGFP ECFP-SSH1ΔC mKeima-Red-Mito-7 pOPTN-EGFP HA-Ubiquitin

Mito Ssh1 Ubiquitin

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Protocol cited in 2 other protocols

Variable analysis

independent variables

pN3-3xFlag-Control
pmRFP-LC3
pMXs-puro GFP-Sqstm1/p62
pBABE-puro mCherry-EGFP-LC3B
pBABE-EGFP
ECFP-SSH1ΔC
EGFP-SSH1
mKeima-Red-Mito-7
pOPTN-EGFP
pMXs-puro GFP-Sqstm1/p62ΔC
pMXs-puro GFP-Sqstm1/p62 D337, 338, 339A (GFP-SQSTM1-LIR)
HA-Ubiquitin
pDR125
pN3-3xFlag-SSH1
mCherry-EGFP-Sqstm1/p62
mCherry-EGFP-Sqstm1^S403A
mCherry-EGFP-Sqstm1^S403E
pN3-Flag-SSH1ΔC
pN3-Flag-SSH1ΔN
pN3-Flag-SSH1ΔN^C393S

dependent variables

Not explicitly mentioned

control variables

pN3-3xFlag-Control

positive controls

Not specified

negative controls

Not specified

Annotations

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