94Q S 97 66 55Q S 19Q S W1W2 97 66 55Q 19Q TAP eluate 94Q S-1 W2 W1 W2 S-1A. Verification of TAP by Western blot (anti-FLAG). S: supernatant of cell lysate.

Slides:



Advertisements
Similar presentations
Supplementary Figures
Advertisements

Supplementary Figure S1 Supplementary Figure S1. Regulation of localization or stability of Cdc25C by ASK1. Effect of ASK1 expression on the stability.
ß-gal NME2 NME1 Actin NME1 NME2 Scrambled RISC Free Supplementary data Figure 1B. Antibody L-15 is specific for NME2 and sc-465 is specific for NME1. Western.
SUPPLEMENTARY MATERIAL consist of two figures: Figure S1 contains two SDS-PAGE gels showing the purification profile of the recombinant yeast and slime.
PreimmuneMonoclonalanti-Aur-AAffinity Purified anti-Aur-A Blot: retic lysate translation product oocyte extract retic lysate translation.
Figure S1. Production of recombinant NS1 protein
RASSF2DSARAH-GF (b) MST1-F (a) M F G 70 kD a 55 kD b 36 kD
1&3: hGH-sc(Fc)2 2&4: hGH-Fc anti-Fc anti-hGH
B A P-AXL P-AXL AXL AXL P-MET P-MET MET MET P-Akt-473 P-Akt-473 AKT
Neal et al.Supplementary Figure S1
Volume 9, Issue 5, Pages (November 1998)
Volume 66, Issue 4, Pages (October 2004)
BRCA1 Is Associated with a Human SWI/SNF-Related Complex
(A) Prdx1−/−MEFs and Prdx1+/+MEFs were stimulated with H2O2 as indicated. (A) Prdx1−/−MEFs and Prdx1+/+MEFs were stimulated with H2O2 as indicated. Protein.
Analysis of CLIP‐170 phosphorylation.
Volume 54, Issue 4, Pages (May 2014)
Knock-down of endogenous PINK1 abrogates Parkin Ser65 phosphorylation.
Volume 5, Issue 5, Pages (May 2004)
The Smc5/6 Complex Is an ATP-Dependent Intermolecular DNA Linker
Hours after rec PR-Set7 release Ladder kDa 15 PR-Set7 Coomassie
by Xingwei Sui, Sanford B. Krantz, and Zhizhuang Zhao
Volume 22, Issue 3, Pages (May 2006)
Rapid ubiquitination of Syk following GPVI activation in platelets
Volume 22, Issue 5, Pages (May 2012)
Volume 23, Issue 1, Pages (July 2006)
(a) (b) FhGALE M U I S W1 W2 E1 E2 E3 M - +
Volume 11, Issue 8, Pages (May 2015)
A A B C D Fraction number b MCF7 lysate fraction A B C D A B C D GST-p851 GST Kinase assay GST-p851.
Volume 9, Issue 5, Pages (May 2016)
Volume 21, Issue 1, Pages (January 2006)
The Polycomb Protein Pc2 Is a SUMO E3
Supplementary Figure 1 RT-PCR BclxL mismatch sc35 Dox BclxS
TopBP1 Controls BLM Protein Level to Maintain Genome Stability
Volume 11, Issue 1, Pages (January 2003)
Lipotransin Molecular Cell
Identification of novel IgGs in alpaca serum.
MED12 interacts with TDRD3 in a CARM1-dependent fashion.
Volume 15, Issue 2, Pages (August 2008)
Volume 123, Issue 2, Pages (October 2005)
Volume 56, Issue 5, Pages (December 2014)
Volume 23, Issue 4, Pages (October 2012)
GSK-3-Mediated Phosphorylation Enhances Maf-Transforming Activity
Volume 51, Issue 6, Pages (September 2013)
Ly6d-L, a Cell Surface Ligand for Mouse Ly6d
Biochemical Characterization of S100A2 in Human Keratinocytes: Subcellular Localization, Dimerization, and Oxidative Cross-Linking1  Rohini Deshpande,
Cyclin G Recruits PP2A to Dephosphorylate Mdm2
Volume 52, Issue 2, Pages (October 2013)
pY-Tie2 (IP with anti-Tie2) Tie2 P-RET RET P-KIT KIT GAPDH P-AXL AXL
Fan Yang, Huafeng Zhang, Yide Mei, Mian Wu  Molecular Cell 
Purified proteins used for in vitro assay.
ONC201 activates the ATF4 pathway through the eIF2α kinases HRI and PKR. ONC201 activates the ATF4 pathway through the eIF2α kinases HRI and PKR. (A) Western.
Figure 4 DNM1 mutations affect protein levels and self-dimerization (A) HeLa cells were transfected with green fluorescent protein (GFP)-tagged mutant.
USP15 Negatively Regulates Nrf2 through Deubiquitination of Keap1
SYCE2 potentiates the steady-state ATM activity.
SLK/LOK-phosphorylated and activated ezrin prevents MISP localization at the cell cortex. SLK/LOK-phosphorylated and activated ezrin prevents MISP localization.
Volume 60, Issue 6, Pages (December 2001)
Volume 34, Issue 5, Pages (June 2009)
SIRT1 Regulates the Function of the Nijmegen Breakage Syndrome Protein
p65 depletion increases basal protein aggregation and insolubilization
Volume 4, Issue 4, Pages (October 1999)
4E-BP is present in an 80 kDa complex in unfertilized eggs.
Effects of visfatin on the cell proliferation and phosphorylation of ERK, Akt, and GSK-3β proteins in HCC cells. Effects of visfatin on the cell proliferation.
Ectopic expression of CA RSK1 mutant protects melanoma cells from PD98059-mediated apoptosis. Ectopic expression of CA RSK1 mutant protects melanoma cells.
Volume 22, Issue 3, Pages (May 2006)
Phosphorylation of the pro-apoptotic protein BAD on serine 155, a novel site, contributes to cell survival  K. Virdee, P.A. Parone, A.M. Tolkovsky  Current.
Volume 41, Issue 4, Pages (February 2011)
Fig. 6 The C9ORF72/SMCR8 complex regulates ULK1.
Volume 123, Issue 2, Pages (October 2005)
RA-9 induces G2–M cell-cycle arrest and caspase-mediated apoptosis in ovarian cancer cells. RA-9 induces G2–M cell-cycle arrest and caspase-mediated apoptosis.
Volume 113, Issue 4, Pages (May 2003)
Presentation transcript:

94Q S Q S 19Q S W1W Q 19Q TAP eluate 94Q S-1 W2 W1 W2 S-1A. Verification of TAP by Western blot (anti-FLAG). S: supernatant of cell lysate loaded to TAP Strep column; W1: sample collected from the flow-through of the Strep column during the washing step; W2: sample collected from the flow-through of the FLAG column during the washing step. A Supplementary Figures

S-1 SYPRO Ruby M 94Q M 19Q 55Q V mHtt wtHtt mHtt S-1B. SDS-PAGE separation of SF- Htt fusion proteins and their associated proteins. After elution of the Htt protein from the Strep and FLAG columns, the proteins that were associated with them was analyzed on a SDS-PAGE gel and stained with a SYPRO Ruby. M, protein molecular weight markers; size, in kD, is indicated on the left. Arrows indicate the bait proteins, which were verified by mass spectrometry. B

* * * S-2 S-2. Prevention of phosphorylation at Ser431 and S432 alters the toxicity of mHtt. HEK293 cells were transfected with indicated plasmid constructs and after 42 hours, the cells were treated with 200µM H 2 O 2 for 24 hour. The percentage (%) of cell viability was measured by the MTT method. * p < n = 4.

Actin 45kD Insoluble Htt (Stacking gel) Soluble Htt (Resolving gel) 95kD 94Q 94QS431 S/A S/D 94QS432 S/D S/A A S-3 S-3A. Prevention of phosphorylation at Ser431 and S432 alters mHtt aggregation. HEK293 cells were transfected with indicated plasmids and after 24 hours of transfection, the cells were collected and subjected Western blot using the antibodies as indicated on the right side of each panel.

* * * * B C S-3B. Quantification of the Western blot result showing protein levels of insoluble (mHtt aggregates found in stacking gel). * P < n = 3. S-3C. Quantification of the Western blot result showing protein levels of soluble mHtt (in resolving gel) (right). * P < n = 3. S-3