Project PXD013143

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Summary

Title

CCT3 acts as upstream of YAP and TFCP2 to be a potential target and tumor biomarker in liver cancer

Description

Recently, we've reported that pro-tumorigenic roles of YAP in liver cancer are indispensable of TFCP2 act as upstream to simultaneously control YAP.The aim of this study is to uncover such proteins and evaluate whether the candidates are potential YAP-associated therapeutic target and diagnostic marker Via mass spectrometry, CCT3 was identified to co-interact with YAP and TFCP2, and importantly.

Sample Processing Protocol

First, the overexpression plasmid of YAP and TFCP2 was used to infect hepatocarcinoma cells. After 24 hours, the protein was collected, washed for 30 min on ice, centrifuged at 13000 rpm for 15 min, protein A / G was used for immunoprecipitation, 4 ℃ shaker overnight, Liquid wash 4 times, multiplate the sample buffer 100 degrees cook 10min, with 1.0 glue run western blot, when the gel ran to half, cut, stained with Coomassie dyeing, and then bleaching, the gel should be cut into small pieces. Desalted dehydration and then air-dried, adding protein precipitation before add to column analysis.

Data Processing Protocol

LC system (Nano Pump, Ultimate 3000, Dionex, Thermofisher) coupled with an ESI-Q-TOFmass spectrometer (maXis, Impact, Bruker Daltonik,Germany).The proteins that were precipitated by YAP and TFCP2 were analyzed by matching software, and the scores were compared. The common proteins were found from the three replicates and the proteins in the blank samples were removed. That is, the rest of the available protein, access to literature, find out transcription factor proteins.

Contact

YA LIU, +8618817567010
Fenyong Sun, Department of Clinical Laboratory, Shanghai Tenth People's Hospital of Tongji University, Shanghai, 200072, China ( lab head )

Submission Date

19/03/2019

Publication Date

11/09/2019

Tissue

liver

Cell Type

hepatocyte

Disease

liver cancer

Modification

Carbamidomethyl
Oxidation

Quantification

Not available

Experiment Type

Gel-based experiment

Assay count

9

Publication

    Liu Y, Zhang X, Lin J, Chen Y, Qiao Y, Guo S, Yang Y, Zhu G, Pan Q, Wang J, Sun F. CCT3 acts upstream of YAP and TFCP2 as a potential target and tumour biomarker in liver cancer. Cell Death Dis. 2019 10(9):644 PubMed: 31501420

Assay

Showing 1 - 9 of 9 results
# Accession Title Proteins Peptides Unique Peptides Spectra Identified Spectra View in Reactome
1 106501 F010238-T1_RB7_01_5880.mzid 636 6438 2545 25397 0
2 106500 F010231-T2_RB8_01_5882.mzid 666 8392 2729 27997 0
3 106499 F010232-T3_RC1_01_5884.mzid 712 10024 3167 27577 0
4 106498 F012210-Y2_RB5_01_5876.mzid 440 5947 1582 26914 0
5 106505 F010239-Y1_RB4_01_5892.mzid 668 8990 3172 17934 0
6 106504 F010233-Y3_RB6_01_5896.mzid 589 9226 2681 27519 0
7 106503 F010468-B1_RD3_01_6235.mzid 60 1393 180 26559 0
8 106502 F010469-B2_RD4_01_6237.mzid 57 920 145 28751 0
9 106506 F010470-B3_RD5_01_6239.mzid 57 1717 152 22985 0