Antibodies |
Mouse monoclonal anti-SRPK2 |
BD Biosciences |
Cat#BD611118; RRID: AB_398429 |
Rabbit polyclonal anti-ACLY |
Cell Signaling Technology |
Cat#4332; RRID: AB_2223744 |
Rabbit monoclonal anti-FASN |
Cell Signaling Technology |
Cat#3180; RRID: AB_2100796 |
Mouse monoclonal anti-S6 |
Cell Signaling Technology |
Cat#2317; RRID: AB_2238583 |
Rabbit monoclonal anti-pS6(S240/S244) |
Cell Signaling Technology |
Cat#5364; RRID: AB_10694233 |
Rabbit polyclonal anti-SCD1 |
Cell Signaling Technology |
Cat#2438; RRID: AB_823634 |
Rabbit polyclonal anti-CHREBP |
Novus |
Cat# NB400-135;RRID:AB_10002435 |
Rabbit polyclonal anti-FAM120A |
GeneTex |
Cat#GTX120824; RRID:AB_10723014 |
Rabbit polyclonal anti-FAM120A |
Thermo Fisher Scientific |
Cat# A303-889A; RRID:AB_2620239) |
Rabbit polyclonal anti-LXR alpha |
Thermo Fisher Scientific |
Cat#PA1-330;RRID:AB_325600 |
Mouse monoclonal anti-SREBP2 |
BD Biosciences |
Cat#557037; RRID:AB_396560 |
Rabbit polyclonal anti-LXR beta |
Thermo Fisher Scientific |
Cat#PA1-333;RRID:AB_2154770 |
Rabbit polyclonal anti-PPAR alpha |
Thermo Fisher Scientific |
Cat#PA1-822A; RRID:AB_2165595 |
Rabbit polyclonal anti-PPAR gamma |
Abcam |
Cat#ab209350; RRID:AB_2890099 |
Rabbit monoclonal anti-p300 |
Cell Signaling Technology |
Cat#54062; RRID:AB_2799450 |
Rabbit monoclonal anti-XBP-1s |
Cell Signaling Technology |
Cat#12782;RRID:AB_2687943 |
Mouse monoclonal anti-GAPDH |
Sigma-Aldrich |
Cat#G8795; RRID: AB_1078991 |
Rabbit polyclonal anti-ACSS2 |
Sigma-Aldrich |
Cat#HPA004141; RRID: AB_1078094 |
Mouse monoclonal anti-V5 |
Sigma-Aldrich |
Cat#V8012; RRID:AB_261888 |
Mouse monoclonal anti-Vinculin |
Sigma-Aldrich |
Cat#V9264; RRID: AB_10603627 |
Rabbit polyclonal anti-RNA polymerase II |
Abcam |
Cat#ab26721; RRID:AB_777726 |
Rabbit monoclonal anti-FDFT1 |
Abcam |
Cat#ab109723; RRID: AB_10859772 |
Mouse monoclonal anti-U1–70k |
Santa Cruz Biotechnology |
Cat#sc-390899; RRID:AB_2801569 |
Rabbit polyclonal anti-SREBP1 |
Santa Cruz Biotechnology |
Cat#sc-8984; RRID: AB_2194223 |
Mouse monoclonal anti-SRSF1 |
Santa Cruz Biotechnology |
Cat#sc-33652; RRID: AB_628248 |
Mouse monoclonal anti-T7 |
EMD Millipore |
Cat#69522; RRID:AB_11211744 |
Mouse monoclonal anti-Phosphoepitope SR proteins |
EMD Millipore |
Cat#MABE50;RRID:AB_10807429 |
Rabbit Normal IgG |
Cell Signaling Technology |
Cat#2729, RRID:AB_1031062 |
Mouse Normal IgG |
Thermo Scientific
|
Cat#10-400-C; RRID:AB_2532980 |
IRDye 800CW Donkey anti-Rabbit IgG |
LI-COR Biosciences |
Cat# 926-32213; RRID:AB_621848 |
IRDye 680RD Donkey anti-Mouse IgG |
LI-COR Biosciences |
Cat# 926-68072; RRID:AB_10953628 |
IRDye 680RD Detection Reagent |
LI-COR Biosciences |
Cat#:926-69100 |
Bacterial and virus strains |
E. coli BL21 |
New England Biolabs |
Cat#C2527 |
Chemicals, peptides, and recombinant proteins |
Rapamycin |
Calbiochem |
Cat#553210 |
Torin1 |
Tocris Bioscience |
Cat#4247 |
Actinomycin D |
Sigma-Aldrich |
Cat#A1410 |
PhosSTOP |
Roche |
Cat#04906837001 |
RNase inhibitor |
Invitrogen |
Cat#10777019 |
RNase T1 |
Thermo Fisher Scientific |
Cat#AM2283 |
RNase A |
Thermo Fisher Scientific |
Cat#12091-021 |
Anti-V5 agarose affinity gel |
Sigma-Aldrich |
Cat#A7345 |
Anti-Flag agarose affinity gel |
Sigma-Aldrich |
Cat# A2220 |
Lipofectamine™ RNAiMAX Transfection Reagent |
Invitrogen |
Cat#13778075 |
DNase I |
Sigma-Aldrich |
Cat#AMPD1 |
TRIzol |
Invitrogen |
Cat#15596018 |
Protein A/G magnetic beads |
Thermo Fisher Scientific |
Cat#88802 |
Protein A/G PLUS-Agarose |
Santa Cruz Biotechnology |
Cat#sc-2003 |
Proteinase K |
New England Biolabs |
Cat#P8107 |
Lipofectamine 2000 reagent |
Invitrogen |
Cat#11668500 |
Phenol:Chloroform:IAA, 25:24:1, pH 6.6 |
Thermo Fisher Scientific |
Cat#AM9730 |
Insulin solution human |
Sigma-Aldrich |
Cat#I9278 |
Crystal Violet |
Sigma-Aldrich |
Cat#C3886 |
SRPIN340 |
Selleck Chemicals |
Cat#S7270 |
SRPKIN-1 |
MedChem Express |
Cat#HY-116856 |
Lipoprotein, low density from human plasma |
Sigma-Aldrich |
Cat#L7914 |
Oleic acid-albumin |
Sigma-Aldrich |
Cat#O3008 |
Fatty acid-free albumin |
Sigma-Aldrich |
Cat#A8806 |
Lipoprotein-deficient serum |
Alfa Aesar |
Cat#BT-907 |
Sodium palmitate |
Sigma-Aldrich |
Cat#P9767 |
Methanol-free 10% formaldehyde |
Polysciences |
Cat#04018-1 |
25-Hydroxycholesterol |
MedChem Express |
Cat#HY-113134 |
Critical commercial assays |
PureLink RNA Mini kit |
Ambion |
Cat#12183018A |
iScript cDNA synthesis kit |
Bio-rad |
Cat#170–8891BUN |
Illumina® Stranded mRNA Prep Kit |
Illumina |
Cat#20040534 |
Gateway LR clonase II enzyme mix |
Invitrogen |
Cat#11791 |
QuickChange site-directed mutagenesis kit |
Strategene |
Cat#200521 |
RNA Clean & Concentrator |
Zymo Research |
Cat#R1016 |
Illumina® Stranded Total RNA Prep, Ligation with Ribo-Zero Plus |
Illumina |
Cat#20040529 |
LightSwitch dual assay kit (promoter assay) |
Active Motif |
Cat#32035 |
Chromatin IP DNA purification kit |
Active Motif |
Cat#58002 |
SYBR Green PCR Master Mix |
Life Technologies |
Cat#4312704 |
QuikChange II XL Site-Directed Mutagenesis Kit |
Strategene |
Cat#200521 |
ChIP-IT Express Kit |
Active Motif |
Cat#53009 |
Deposited data |
Raw RNAseq data (siNTC and siFAM120A) |
This paper |
GSE207172 |
Raw RNAseq data (siNTC, siFAM120, and siSRSF1) |
This paper |
GSE229657 |
Raw data of Images and WB |
This paper, Mendeley Data |
DOI: 10.17632/rgfgx9xb8w.1
|
Experimental models: Cell lines |
Human: Renal angiomyolipoma-derived LAM 621–101 (TSC2−/−) |
Drs. Jane Yu and Elizabeth Henske |
Yu et al., 2004; RRID: CVCL_S879 |
Human: HEK293E |
ATCC |
Cat#293 c18; RRID: CVCL_6974 |
Human: HEK293T |
GenHunter |
Cat#Q401; RRID: CVCL_0063 |
Human: MCF7 |
ATCC |
Cat#HTB-22; RRID: CVCL_0031 |
Human: DLD1 |
ATCC |
Cat#CCL-221; RRID: CVCL_0248 |
Human: NCI-H1299 |
ATCC |
Cat#CRL-5803; RRID: CVCL_0060 |
Human: LNCaP |
ATCC |
Cat#CRL-1740; RRID:CVCL_1379 |
Experimental models: Organisms/strains |
nu/nu athymic mice |
Envigo |
Cat#069 |
Oligonucleotides |
siRNA negative control-#1(siNTC) |
Sigma-Aldrich |
SIC001 |
siRNA negative control-#2(siNTC) |
Sigma-Aldrich |
SIC002 |
siRNA (Human SRSF1) |
Sigma-Aldrich |
SASI_Hs01_00115059 |
siRNA (Human SRSF1) |
Sigma-Aldrich |
SASI_Hs02_00342609 |
siRNA (Human UPF1) |
Sigma-Aldrich |
SASI_Hs01_00101017 |
siRNA (Human UPF1) |
Sigma-Aldrich |
SASI_Hs01_00101018 |
siRNA (Human U1–70k/SNRNP70) |
Sigma-Aldrich |
SASI_Hs02_00335028 |
siRNA (Human U1–70k/SNRNP70) |
Sigma-Aldrich |
SASI_Hs01_00165101 |
siRNA (Human FAM120A) |
Sigma-Aldrich |
SASI_Hs01_00149752 |
siRNA (Human FAM120A) |
Sigma-Aldrich |
SASI_Hs02_00345898 |
siRNA (Human FAM120A) |
Sigma-Aldrich |
SASI_Hs02_00345899 |
siRNA (Human SREBP1) |
Sigma-Aldrich |
SASI_Hs01_00051828 |
siRNA (Human SREBP1) |
Sigma-Aldrich |
SASI_Hs01_00051829 |
siRNA (Human SREBP1) |
Sigma-Aldrich |
SASI_Hs01_00051830 |
siRNA (Human SREBP2) |
Sigma-Aldrich |
SASI_Hs01_00075424 |
siRNA (Human SREBP2) |
Sigma-Aldrich |
SASI_Hs01_00075425 |
siRNA (Human SREBP2) |
Sigma-Aldrich |
SASI_Hs01_00075426 |
Primers for qPCR analysis, see Table S2
|
This paper |
N/A |
Recombinant DNA |
ACSS2 promoter construct |
SwitchGear Genomics |
Cat#S715263 |
SCD1 promoter construct |
SwitchGear Genomics |
Cat#S714484 |
pLX304-SRSF1-V5 |
Harvard plasmid |
Cat#HsCD00420441 |
Lentiviral packaging and envelope plasmids |
Dr. David Baltimore |
N/A |
pcDNA3-EGFP |
Addgene |
Cat#13031 |
pKH3-HA-SRPK2-WT, K110M |
Dr. John Blenis |
Lee et al., 2007 |
pLNCX-HA-SRPK2-S494A/S497A |
Dr. John Blenis |
Lee et al., 2007 |
pLX304-FAM120A-V5 |
Harvard plasmid |
HsCD00440255 |
pENTR1A-Flag-FAM120A(WT) |
This paper |
N/A |
pENTR1A-Flag-FAM120A(∆RBD) |
This paper |
N/A |
pLenti-CMV-Puro-DEST |
Addgene |
Cat#17452 |
pENTR223-SRSF1 |
Harvard plasmid |
Cat#HsCD00367645 |
pLenti-CMV-Puro-SRSF1 |
This paper |
N/A |
pLenti-CMV-Puro-FAM120A |
This paper |
N/A |
pLenti-CMV-Puro-Flag-FAM120A(WT) |
This paper |
N/A |
pLenti-CMV-Puro-Flag-FAM120A(∆RBD) |
This paper |
N/A |
pENTR223-FAM120A |
Harvard plasmid |
Cat#HsCD00438604 |
T7-SRSF1(WT) |
Dr. Adrian Krainer |
Fregoso et al., 2013 |
T7-SRSF1(∆RRM1) |
Dr. Adrian Krainer |
Fregoso et al., 2013 |
T7-SRSF1(∆RRM2) |
Dr. Adrian Krainer |
Fregoso et al., 2013 |
T7-SRSF1(∆RS) |
Dr. Adrian Krainer |
Fregoso et al., 2013 |
pLKO.1-puro-Non-Mammalian shRNA Control |
Sigma-Aldrich |
SHC002 |
pLKO.1-puro-shSRPK2 |
Sigma-Aldrich |
TRCN0000006274 |
pLKO.1-puro-shFAM120A |
Sigma-Aldrich |
TRCN0000074918 |
pLKO.1-puro-shSREBP1 |
Sigma-Aldrich |
TRCN0000055325 |
pLKO.1-puro-shSREBP2 |
Sigma-Aldrich |
TRCN0000020667 |
Software and algorithms |
IRFinder-S |
Lorenzi et. al., 2021
|
https://europepmc.org/article/med/34749764
|
HISAT2 v2.2.1 |
Kim et. al., 2019
|
https://www.nature.com/articles/s41587-019-0201-4
|
StringTie2 v2.1.4 |
Kovaka et. al., 2019
|
https://genomebiology.biomedcentral.com/articles/10.1186/s13059-019-1910-1
|
DESeq2 (v1.30.1) |
Love et. al., 2014
|
https://genomebiology.biomedcentral.com/articles/10.1186/s13059-014-0550-8
|
Enhanced Volcano v1.8.0 R package |
Blighe, 2022
|
https://bioconductor.org/packages/devel/bioc/vignettes/EnhancedVolcano/inst/doc/EnhancedVolcano.html
|
Adobe Photoshop |
Adobe |
Adobe |
Odyssey imaging system |
LI-COR Biosciences |
LI-COR Biosciences |
enrichR v3.0 package |
Webb et al., 2014 |
https://git.ecdf.ed.ac.uk/sgrannem/pycrac
|
EI-MAVEN, Version 0.12.0 |
Agrawal et al, 2019
|
Elucidata |
VAST-tools |
Irimia et. al., 2014
|
https://www.sciencedirect.com/science/article/pii/S0092867414015128?via%3Dihub
|
BioRender |
BioRender |
https://www.biorender.com/
|