|
| Antibodies |
|
|
|
| Mouse monoclonal SCoR/AKR1A1 |
Santa Cruz Biotechnology |
sc-100500 RRID:AB_118799 |
| Rabbit polyclonal GAPDH |
Proteintech |
10494-1-AP RRID:AB_2263076 |
| Rabbit monoclonal LDLR |
Abcam |
ab52818 RRID:AB_881213 |
| Goat polyclonal mouse PCSK9 |
R&D Systems |
AF3985 RRID:AB_2044717 |
| Rabbit monoclonal human PCSK9 |
Cell Signaling |
85,813 RRID:AB_2800064 |
| Rabbit polyclonal PCSK9 |
Proteintech |
55206-1-AP RRID:AB_10918272 |
| Rabbit monoclonal HMGCR |
Abcam |
ab174830 RRID:AB_2749818 |
| Rabbit polyclonal ACAT2 |
Cell Signaling |
11,814 |
| Rabbit polyclonal ARH |
Proteintech |
13213-1-AP RRID:AB_2135283 |
| Rabbit monoclonal SAR1A/B |
Abcam |
ab155278 |
| Rabbit polyclonal SEC23A |
Cell Signaling |
8162 RRID:AB_10859891 |
| Goat polyclonal SEC23A |
Novus Biologicals |
NBP1-47201 RRID:AB_10010164 |
| Rabbit polyclonal SEC24A |
Proteintech |
15958-1-AP RRID:AB_2186118 |
| Rabbit polyclonal SURF4 |
Sigma-Aldrich |
SAB2108065 |
| Rabbit monoclonal V5 |
Cell Signaling |
13,202 RRID:AB_2687461 |
| Mouse monoclonal calnexin |
Proteintech |
66903-1-Ig RRID:AB_2882231 |
| Goat polyclonal rabbit IgG Alexa Fluor 488 |
Thermo Fisher |
A-11008 RRID:AB_143165 |
| Goat polyclonal mouse IgG Alexa Fluor 594 |
Thermo Fisher |
A-11005 RRID:AB_2534073 |
| Donkey polyclonal rabbit IgG Alexa Fluor 488 |
Thermo Fisher |
A-32790 RRID:AB_2762833 |
| Donkey polyclonal goat IgG Alexa Fluor 594 |
Thermo Fisher |
A-11058 RRID:AB_142540 |
|
| Chemicals, peptides, and recombinant proteins |
|
|
|
| Sodium ascorbate |
Sigma-Aldrich |
Cat #11140 |
| S-methyl thiomethansulfonate, MMTS |
Sigma-Aldrich |
Cat #64306 |
| Thiopropyl Sepharose 6B |
GE Lifesciences |
Cat #17-0420-01 |
| Cysteine ethyl ester hydrochloride |
Sigma-Aldrich |
Cat #C121908 |
| Coenzyme A |
Sigma-Aldrich |
Cat #C4780 |
| Cysteine |
Sigma-Aldrich |
Cat #C6852 |
| V5 affinity agarose |
Sigma-Aldrich |
Cat #A7345 |
| V5 peptide |
Sigma-Aldrich |
Cat #V7754 |
| FLAG affinity agarose |
Sigma-Aldrich |
Cat #A2220 |
| FLAG peptide |
Sigma-Aldrich |
Cat #F3290 |
| 3X FLAG peptide |
Sigma-Aldrich |
Cat #F4799 |
| IodoTMT |
Thermo Fisher |
Cat #90101 |
| Iodoacetamide |
Sigma-Aldrich |
Cat #I1149 |
| Anti-TMT resin |
Thermo Fisher |
Cat #90076 |
| AL-1576 |
TCG Lifesciences, WuXi China |
Custom synthesis |
|
| Critical commercial assays |
|
|
|
| High capacity RNA-to-cDNA kit |
Applied Biosystems |
Cat #4387406 |
| TaqMan University Master Mix II (without UNG) |
Thermo Fisher |
Cat #4440043 |
| LDLR TaqMan Probe |
Thermo Fisher |
Cat # Mm01177349_m1 |
| PCSK9 TaqMan Probe |
Thermo Fisher |
Cat # Mm0126310_m1 |
| SREBF2 TaqMan Probe |
Thermo Fisher |
Cat # Mm01306292_m1 |
| GAPDH TaqMan Probe |
Thermo Fisher |
Cat # Mm99999915_g1 |
| Cholesterol Assay |
Abcam |
Cat #ab65359 |
| Triglyceride Assay |
Cayman Chemicals |
Cat #10010303 |
| Mouse PCSK9 ELISA |
R&D Systems |
Cat #MPC900 |
| Mouse alpha-1-antitrypsin ELISA |
Crystal Chem |
Cat #80631 |
|
| Deposited data |
|
|
|
| Blot and image raw files |
Mendeley Data |
https://doi.org/10.17632/z2fm5g6tcf
|
|
| Experimental models: Cell lines |
|
|
|
| Human: HepG2 stably expressing non-targeting shRNA |
This study |
N/A |
| Human: HepG2 stably expressing SCoR-targeting shRNA |
This study |
N/A |
| Human: 293T/17 |
ATCC |
Cat #CRL-11268 RRID:CVCL_1926 |
| Human: SCoR-deficient HEK293 |
Stomberski et al., (2019a)
|
PMID:30538128 |
| Human: SCoR-deficient HEK293 stably expressing PCSK9 |
This study |
N/A |
|
| Experimental models: Organisms/strains |
|
|
|
| Mouse: AKR1A1tm1Dgen
|
Deltagen |
N/A |
| Mouse: C57BL/6J |
The Jackson Laboratory |
Strain #000664 RRID:IMSR_JAX:000,664 |
| Mouse: B6.129S7-Ldlrtm1Her/J |
The Jackson Laboratory |
Strain #002207 RRID:IMSR_JAX:002,207 |
| Mouse: B6.SJL-Tg(APOA-CETP) 1dsgTg(APOB100)1102SgyN10 |
Taconic |
Strain #3716-M RRID:IMSR_TAC:3716 |
| Mouse: B6.129P2-Apoetm1Unc/J |
The Jackson Laboratory |
Strain #002052 RRID:IMSR_JAX:002,052 |
| Mouse: AKR1A1−/−/LDLR−/−
|
This Study |
N/A |
|
| Oligonucleotides |
|
|
|
| Mouse Genotyping Primers: See Table S1
|
This Study |
N/A |
| PCSK9, SURF4, SAR1B mutagenesis primers: See Table S2
|
This Study |
N/A |
| Mammalian non-targeting shRNA |
Sigma-Aldrich |
Cat #SHC002 |
| SCoR-targeting shRNA |
Sigma-Aldrich |
Cat # TRCN0000231969 |
|
| Recombinant DNA |
|
|
|
| pEntr223.1-PCSK9 |
Dharmacon |
Cat #OHS5893-202503467 |
| pEntr223.1-PCSK9-Fusion |
DNASU |
Cat # HsCD00294884 |
| pCSF107mT-GATEWAY-3′-FLAG |
Todd Stukenberg Lab |
Addgene Cat #67619 |
| pcDNA-Dest40 |
Thermo Fisher |
Cat #12274015 |
| pcDNA-Dest40-PCSK9WT
|
This Study |
N/A |
| pcDNA-Dest40-PCSK9C301A
|
This Study |
N/A |
| pCSF107mT-PCSK9-3′-FLAG |
This Study |
N/A |
| pDONR221-SURF4 |
DNASU |
Cat # HsCD00044097 |
| pcDNA3.2/nV5-DEST |
Thermo Fisher |
Cat #12290010 |
| pcDNA3.2/nV5-SURF4WT
|
This Study |
N/A |
| pcDNA3.2/nV5-SURF4C32A
|
This Study |
N/A |
| pCMV6-SAR1BWT
|
Origene |
Cat #RC213692 |
| pCMV6-SAR1BC102A/C178A
|
This Study |
N/A |
| pCMV6-AKR1A1WT
|
Origene |
Cat #RC200302 |
|
| Software and algorithms |
|
|
|
| GraphPad Prism v7 |
GraphPad Inc |
N/A |
|
| Other |
|
|
|
| Mouse Diet: AIN-93M + 1% ascorbic acid |
Research Diets |
Cat #D16021012, custom diet |
| Mouse Diet: AIN-93M + 0.0125% AL-1576 |
Research Diets |
Cat #D15111101, custom diet |
| Mouse Diet: AIN-93M |
Research Diets |
Cat #D10012M |
