AGO1      shRNA in HepG2      Control: NT_BGHLV14_1

General Information
RBPAGO1
Cell_LineHepG2
MethodshRNA
Exp_NameAGO1-BGHLV14-HepG2
ENCODE_series_IDENCSR213SDO
Batch_IDBGHLV14
Pool IDPool-140915
Local_Set_Nameset14
ENCODE_KD_Exp_IDENCSR533HXS
ENCODE_CN_Exp_IDENCSR003EKR
Rep1EIF2C1_BGHLV14-13
Rep2EIF2C1_BGHLV14-14
CN1NT_BGHLV14-1_1
CN2NT_BGHLV14-2_1
Rep1_qPCR86.1
Rep2_qPCR85.1
Rep1_WB93.1
Rep2_WB94.1
Antibody Cat#RN028PW
Antibody Lot#RN028PW
Antibody DCC IDENCAB179WCQ
StatusReleased
ProjectENCODE3
ID389




Experiment Information (Status: Released)
BGHLV14-13BGHLV14-13BGHLV14-14BGHLV14-14
idx107109108110
TRCN#_or_BGC#TRCN0000007860TRCN0000007860TRCN0000007860TRCN0000007860
shRNA_or_gRNA_sequenceGCTGACAAGAATGAGCGAATTGCTGACAAGAATGAGCGAATTGCTGACAAGAATGAGCGAATTGCTGACAAGAATGAGCGAATT
PAM
NameEIF2C1EIF2C1EIF2C1EIF2C1
Sample_IDBGHLV14-13BGHLV14-13BGHLV14-14BGHLV14-14
transduction_Date7/4/20147/4/20147/4/20147/4/2014
days
RBP_nameAGO1AGO1AGO1AGO1
qPCR_result86.186.185.185.1
Ave_qPCR85.685.6
RT-qPCR_primer-FCGCATCATCTTCTACCGAGACGCATCATCTTCTACCGAGA
RT-qPCR_primer-RTTTCCAGTTTGATGCAGGCTTTCCAGTTTGATGCAGGC
protein_conc4355435540684068
WB_result93.186.494.187.8
Ave_WB93.687.1
WB_DONE_date2/25/20162/25/2016
MW97kDa97kDaWESWES
IP11FU'S
antibody_Cat#RN028PW#5053RN028PW#5053
Antibody DCC IDENCAB179WCQENCAB189GDEENCAB189GDE
submitted_to_DCC_date3/8/20163/8/2016
Rep1_TPM2.42.400
Rep2_TPM1.41.400
Action
Library_start_date7/30/20147/30/2014
repeat_library
Note
ID2164216621652167




Western Blot
RBP Antibody InfoPrimary-HepG2Secondary-HepG2Primary-K562Secondary-K562Primary-UBERONPrimary-Hela
AGO1Product_ID: RN028PW
Lot_ID: 2
Source: MBLI
Target Name: AGO1-human
EIF2C1-HEPG2-RN028PW.png<br>Caption: Western blot following shRNA against AGO1/EIF2C1 in HepG2 whole cell lysate using EIF2C1 specific antibody. Lane 1 is a ladder, lane 2 is HepG2 non-targeting control knockdown, lane 3 and 4 are two different shRNAs against EIF2C1. EIF2C1 protein appears as the green arrow, GAPDH serves as a control and appears in red arrow.
K562_MBLI_RN028PW_002_EIF2C1.png<br>Caption: IP-WB characterization of AGO1 specific antibody in K562 cell line . Lane 1 is 2.5% of five million K562 whole cell lysate Input, lane 2 is 2.5% of supernatant after immunoprecipitation and Lane 3 is 50% of IP enrichment using rabbit polyclonal Anti-EIF2C1(AGO1)(Human)pAb. This antibody passes preliminary validation and will be further pursued for primary and secondary validation.
EIF2C1-k562-RN028PW.png<br>Caption: Western blot following shRNA against EIF2C1 in K562 whole cell lysate using EIF2C1 specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different shRNAs against EIF2C1. EIF2C1 protein appears as the green arrow, GAPDH serves as a control and appears in red arrow.
EIF2C1_Secondary_Western.png<br>Caption: Western blot following shRNA against AGO1 in K562 whole cell lysate using AGO1 specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different shRNA against AGO1. AGO1 protein appears as the green arrow, Tubulin serves as a control and appears in red arrow.
AGO1Product_ID: 5053
Lot_ID: 1
Source: Cell Signaling Technology
Target Name: AGO1-human
AGO1_HepG2.png<br>Caption: IP-Western Blot analysis of HepG2 whole cell lysate using AGO1/EIF2C1 specific antibody. Lane 1 is 3% of ten million whole cell lysate input (lane under '15% input') , lane 2 is 20% of IP enrichment using rabbit normal IgG (lane under 'IgG') and lane 3 is 20% IP enrichment using rabbit monoclonal anti-AGO1/EIF2C11 antibody (lanes under 'anti-AGO1'). Asterisk indicates heavy chain of antibody.
EIF2C1-HEPG2-FU%27S.png<br>Caption: Western blot following shRNA against EIF2C1 in HepG2 whole cell lysate using EIF2C1 specific antibody. Lane 1 is a ladder, lane 2 is HepG2 non-targeting control knockdown, lane 3 and 4 are two different shRNAs against EIF2C1. EIF2C1 protein appears as the green arrow, GAPDH serves as a control and appears in red arrow.
AGO1_K562.png<br>Caption: IP-Western Blot analysis of K562 whole cell lysate using AGO1/EIF2C1 specific antibody. Lane 1 is 3% of ten million whole cell lysate input (lane under '15% input') , lane 2 is 20% of IP enrichment using rabbit normal IgG (lane under 'IgG') and lane 3 is 20% IP enrichment using rabbit monoclonal anti-AGO1/EIF2C11 antibody (lanes under 'anti-AGO1'). Asterisk indicates heavy chain of antibody.




Experiment Status
BGHLV14-13BGHLV14-14
Sample_IDBGHLV14-13BGHLV14-14
Sample NameEIF2C1_BGHLV14-13EIF2C1_BGHLV14-14
Sample NameEIF2C1_BGHLV14-13EIF2C1_BGHLV14-14
RBPEIF2C1EIF2C1
Cell_LineHepG2HepG2
Exp UID
StatusReleasedReleased
Status_date2014-11-042014-11-04
ProjectENCODE3ENCODE3
Note
ID500501




Library-Prep Information
BGHLV14-13BGHLV14-14
Sample #
Sample NameAGO1-BGHLV14-13AGO1-BGHLV14-14
Sample_Name_AliasEIF2C1_BGHLV14-13EIF2C1_BGHLV14-14
Index Well PositionS18S19
Index_tableIDT_TruSeqSingleIndex_24IDT_TruSeqSingleIndex_24
LibPrep_date2014-07-302014-07-30
Lib_IDLib-140730Lib-140730
Tecan_Location
Tecan
Tecan_date
Size_bp
Peak_Molarity
libSampleQC_DNA_WellDNA_Library_LV13-14/set1/B9.pngDNA_Library_LV13-14/set1/C9.png
RIN9.69.5
libSample_RNA_WellRNA_Library_LV13-14-16/set1/E5.pngRNA_Library_LV13-14-16/set1/F5.png
SampleQC_methodTapeStationTapeStation
SampleQC_date
Sample_IDBGHLV14-13BGHLV14-14
RBPAGO1AGO1
Batch_IDBGHLV14BGHLV14
WB_result93.10094.100
Library DescriptionTruSeq mRNATruSeq mRNA
Repeat_Library_Suffix
Lib_StatusSendToSequenceSendToSequence
ProjectENCODE3ENCODE3
ID21842185




Sequencing Information
BGHLV14-13BGHLV14-14
Sample_IDBGHLV14-13BGHLV14-14
Sample NameEIF2C1_BGHLV14-13EIF2C1_BGHLV14-14
Pool IDPool-140915Pool-140915
LocalServer_folderset14set14
total_reads51,620,00047,580,000
total_aligned_reads48,983,66244,786,554
unique_aligned_reads46,279,47442,234,552
percent_uniqueAligned0.896540.88765
correlation_replicates0.9960040.996004
spikein_reads234,004196,844
percent_spikeins0.004530.00414
original_ReadLength100100
QC_StatusSubmittedSubmitted
ID826827




Data Submission Information
BGHLV14-13BGHLV14-13
ENCODE_aliasbrenton-graveley:EIF2C1_BGHLV14brenton-graveley:AGO1-BGHLV14-HepG2
ENCODE_accessionENCSR533HXSENCSR213SDO
object_typeexperimentgene_silencing_series
assay_typeknockdown followed by RNA-seqknockdown followed by RNA-seq
Sample_IDBGHLV14-13BGHLV14-13
Sample NameEIF2C1_BGHLV14-13EIF2C1_BGHLV14-13
SelectedYesYes
StatusReleasedReleased
Status_Date2014-10-302022-03-01
ProjectENCODE3ENCODE3
protocol_URL
ID28006583




File Information
file_namefile_pathfile_formatoutput_typeassay_typelabSample IDplatformread_lengthrun_typepaired_endassemblygenome_annotationderived_fromcontrolled_byProjectNoteFILE_ID
EIF2C1_BGHLV14-13_GTCCGC_L001_R1_001.fastq.gzENCODE_DATA/set14/fastqfastqreadsknockdown followed by RNA-seqGraveley Lab826HiSeq2000100paired-ended1NT_BGHLV14-1_1_ACAGTG_L008_R1_001.fastq.gz,NT_BGHLV14-2_1_GCCAAT_L008_R1_001.fastq.gzENCODE313792
EIF2C1_BGHLV14-13_GTCCGC_L001_R2_001.fastq.gzENCODE_DATA/set14/fastqfastqreadsknockdown followed by RNA-seqGraveley Lab826HiSeq2000100paired-ended2NT_BGHLV14-1_1_ACAGTG_L008_R2_001.fastq.gz,NT_BGHLV14-2_1_GCCAAT_L008_R2_001.fastq.gzENCODE313854
EIF2C1_BGHLV14-13_Aligned.sortedByCoord.out.bamENCODE_DATA/set14/STAR_ENCORE2_file_linksbamalignmentsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13_GTCCGC_L001_R1_001.fastq.gz,EIF2C1_BGHLV14-13_GTCCGC_L001_R2_001.fastq.gzENCORE213916
EIF2C1_BGHLV14-13_Signal.Unique.strand+.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig+uniqueplus strand signal of unique readsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13/Aligned.sortedByCoord.out.bamENCORE213978
EIF2C1_BGHLV14-13_Signal.Unique.strand-.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig-uniqueminus strand signal of unique readsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13/Aligned.sortedByCoord.out.bamENCORE214040
EIF2C1_BGHLV14-13_Signal.UniqueMultiple.strand+.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig+allplus strand signal of all readsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13/Aligned.sortedByCoord.out.bamENCORE214102
EIF2C1_BGHLV14-13_Signal.UniqueMultiple.strand-.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig-allminus strand signal of all readsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13/Aligned.sortedByCoord.out.bamENCORE214164
EIF2C1_BGHLV14-13_quant.sfENCODE_DATA/set14/salmon_ENCORE2transcript_quantificationtranscript quantificationsknockdown followed by RNA-seqGraveley Lab826GRCh38V40EIF2C1_BGHLV14-13_GTCCGC_L001_R1_001.fastq.gz,EIF2C1_BGHLV14-13_GTCCGC_L001_R2_001.fastq.gzENCORE214226
EIF2C1_BGHLV14-14_GTGAAA_L001_R1_001.fastq.gzENCODE_DATA/set14/fastqfastqreadsknockdown followed by RNA-seqGraveley Lab827HiSeq2000100paired-ended1NT_BGHLV14-1_1_ACAGTG_L008_R1_001.fastq.gz,NT_BGHLV14-2_1_GCCAAT_L008_R1_001.fastq.gzENCODE313793
EIF2C1_BGHLV14-14_GTGAAA_L001_R2_001.fastq.gzENCODE_DATA/set14/fastqfastqreadsknockdown followed by RNA-seqGraveley Lab827HiSeq2000100paired-ended2NT_BGHLV14-1_1_ACAGTG_L008_R2_001.fastq.gz,NT_BGHLV14-2_1_GCCAAT_L008_R2_001.fastq.gzENCODE313855
EIF2C1_BGHLV14-14_Aligned.sortedByCoord.out.bamENCODE_DATA/set14/STAR_ENCORE2_file_linksbamalignmentsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14_GTGAAA_L001_R1_001.fastq.gz,EIF2C1_BGHLV14-14_GTGAAA_L001_R2_001.fastq.gzENCORE213917
EIF2C1_BGHLV14-14_Signal.Unique.strand+.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig+uniqueplus strand signal of unique readsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14/Aligned.sortedByCoord.out.bamENCORE213979
EIF2C1_BGHLV14-14_Signal.Unique.strand-.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig-uniqueminus strand signal of unique readsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14/Aligned.sortedByCoord.out.bamENCORE214041
EIF2C1_BGHLV14-14_Signal.UniqueMultiple.strand+.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig+allplus strand signal of all readsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14/Aligned.sortedByCoord.out.bamENCORE214103
EIF2C1_BGHLV14-14_Signal.UniqueMultiple.strand-.bwENCODE_DATA/set14/STAR_ENCORE2_file_linksbigWig-allminus strand signal of all readsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14/Aligned.sortedByCoord.out.bamENCORE214165
EIF2C1_BGHLV14-14_quant.sfENCODE_DATA/set14/salmon_ENCORE2transcript_quantificationtranscript quantificationsknockdown followed by RNA-seqGraveley Lab827GRCh38V40EIF2C1_BGHLV14-14_GTGAAA_L001_R1_001.fastq.gz,EIF2C1_BGHLV14-14_GTGAAA_L001_R2_001.fastq.gzENCORE214227