KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
|
| ||
| Rabbit anti-SREBP-1 | Rong et al.33 | 20B12 |
| Rabbit anti-SREBP-2 | Rong et al.33 | 22D5 |
| Mouse anti-Insig | Jo et al.55 | 17H1 |
| Rabbit anti-Calnexin | Enzo | Cat#ADI-SPA-860-F |
| Rabbit anti-β-actin | Cell Signaling | Cat#4970S |
| Rabbit anti-LSD1 | Cell Signaling | Cat#2184 |
| Rabbit anti-SCD-1 | Cell signal | Cat#2794 |
| Mouse anti-CREB | ThermoFisher | Cat#35-0900 |
| Mouse anti-EEA1 | Sigma-Aldrich | Cat#E7659 |
| Mouse anti-Cytochrome C | Sigma-Aldrich | Cat#MAB1800 |
| Mouse anti-NPC-2 | Sigma-Aldrich | Cat#MABS440 |
| Mouse anti-GM130 | BD Biosciences | Cat#610822 |
| Mouse anti-Prohibitin-1 | Santa Cruz | Cat#sc-56467 |
| Rabbit anti-LAMP1 | Abcam | Cat#ab24170 |
| Rabbit anti-α-tubulin | Abcam | Cat#ab6160 |
| Mouse anti-PMP70 | Sigma-Aldrich | Cat#SAB4200181 |
| Mouse anti-His | Sigma-Aldrich | Cat#05-949 |
| Rabbit anti-NaK ATPase | Abcam | Cat#ab76020 |
| Rabbit anti-HA | Cell Signaling | Cat#3724 |
| Rabbit anti-mCherry | Abcam | Cat#ab167453 |
| Rabbit anti-ERGIC53 (LMAN-1) | Abcam | Cat#ab125006 |
| Goat Anti-Rabbit IgG, F(ab’)2 | Jackson Immuno | Cat#111-035-047 |
| Goat Anti-Mouse IgG, F(ab’)2 | Jackson Immuno | Cat#115-035-072 |
| Li-COR IRDY 800 goat anti-Rabbit IgG | Fisher Scientific | Cat# NC9401842 |
| Li-COR IRDY 680 goat anti-Mouse IgG | Fisher Scientific | Cat# NC0252290 |
|
| ||
| Bacterial and virus strains | ||
|
| ||
| pAAVscCB6 | Gene Therapy Center at University of Massachusetts | Plasmid#P1023G10 |
| AAV-DJ-eGFP | VectorBiolabs | Cat#7118 |
| pAAV-CAG-shuttle-WPRE-GFP | Applied Viromics | Plasmid#0916 |
| AAV-DJ Helper Free Expression System | Cell Biolabs | VPK-410-DJ |
| pAAV8 | Addgene | Plasmid#112864 |
| pX459 v2.0 | Addgene | Plasmid#134451 |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| DGAT2 inhibitor (PF-06424439) | Sigma-Aldrich | Cat# PZ0233 |
| Tyloxapol (triton WR-1339) | Sigma-Aldrich | Cat#T8761 |
| HPCD | ThermoFisher | Cat#NC0999732 |
| His-tagged ALOD4 | Infante and Radhakrishnan40 | N/A |
| PC | Avanti | Cat#840052C |
| PE | Avanti | Cat#840021C |
| PI | Avanti | Cat#840042C |
| PS | Avanti | Cat#840032C |
| Ethanolamine | Sigma-Aldrich | Cat#398136 |
| Palmitic acid | Sigma-Aldrich | Cat#P5585 |
| Liberase | Sigma-Aldrich | Cat#5401127001 |
| Dnase1 | Sigma-Aldrich | Cat# 10104159001 |
| HBSS, − Ca2+, − Mg2+, no phenol red | ThermoFisher | Cat#14175095 |
| HBSS, + Ca2+, + Mg2+, no phenol red | ThermoFisher | Cat#14025092 |
| Percoll | Sigma-Aldrich | Cat# P1644 |
| OptiPrep™ Density gradient medium stock | Sigma-Aldrich | Cat# D1556 |
| Flavopiridol | Sigma-Aldrichh | Cat# F3055 |
| eBioscience™ 1× RBC lysis buffer | ThermoFisher | Cat#00-4333-57 |
| ATP | Sigma-Aldrich | Cat#10519987001 |
| GTP | Sigma-Aldrich | Cat#10106399001 |
| Creatine kinase | Sigma-Aldrich | Cat#10127566001 |
| Creatine phosphate | Sigma-Aldrich | Cat#10621714001 |
| Fat free diet | MP biomedicals | Cat# 960238 |
|
| ||
| Critical commercial assays | ||
|
| ||
| RNA STAT-60 kit | TEL TEST | Cat#NC9489785 |
| DNA-free, DNA removal kit | Invitrogen | Cat#1906 |
| Taqman reverse transcription reagents | Applied Biosystems | Cat#N8080234 |
| 2× SYBR Green PCR master Mix | Applied Biosystems | Cat#4309155 |
| Roche protease inhibitor | Rocke | Cat#1836170 |
| SuperSignal West Pico Chemiluminescent Substrate | ThermoFisher | Cat#34580 |
| 10× Genomics Chromium Single Cell 3’ Library and Gel Beads Kit (version 3) | 10× Genomics | Cat#1000075 |
| Pierce BCA Protein Assay Kit | ThermoFisher | Cat#23225 |
|
| ||
| Deposited data | ||
|
| ||
| All single cell sequencing raw data | GEO repository | GSE250338 |
| Uncropped high-resolution blots | Data S1-Source Data | N/A |
| Values used to create all graphs | Data S1-Source Data | N/A |
|
| ||
| Experimental models: Cell lines | ||
|
| ||
| Hepa-1c1c7 | ATCC | Cat#CRL-2026 |
| SV589 | Human Genetic Cell Repository | Cat#GM639 |
| HEK293S GnTI | ATCC | Cat#CRL-3022 |
|
| ||
| Experimental models: Organisms/strains | ||
|
| ||
| C57Bl/6J mouse | Jackson Laboratory | Stock#000664 |
| ob/ob mouse | Jackson Laboratory | Stock#000632 |
| Cas9 knockin mouse | Jackson Laboratory | Stock#026179 |
| TghSREBP-1c rat | Owen et al.36 | N/A |
|
| ||
| Oligonucleotides | ||
|
| ||
| 5’-caccGCCAGAAGTGGTCGTGCAATC-3’ | N/A | OL1243 |
| 5’-aaacGATTGCACGACCACTTCTGGC-3’ | N/A | OL1244 |
| 5’-caccGAGCCTCGTGCTCTTCTCGGT-3’ | N/A | OL1247 |
| 5’-aaacACCGAGAAGAGCACGAGGCTC-3’ | N/A | OL1248 |
| 5’-caccGTCACCTGGGCCCAAAAAGTG-3’ | N/A | OL1515 |
| 5’-aaacCACTTTTTGGGCCCAGGTGAC-3’ | N/A | OL1516 |
| 5’-caccGATGCTTGCAATCACATCAGG-3’ | N/A | OL1517 |
| 5’-aaacCCTGATGTGATTGCAAGCATC-3’ | N/A | OL1518 |
| 5’-accGGATCTGCCCTGTCACGCGAG-3’ | N/A | Dgat2sg-F |
| 5’-aacCTCGCGTGACAGGGCAGATCC-3’ | N/A | Dgat2sg-R |
| 5’-accGCTGAGGATACGGCACTGGCG-3’ | N/A | Insig-1sg-F |
| 5’-aacCGCCAGTGCCGTATCCTCAGC-3’ | N/A | Insig-1sg-R |
| 5’-accGTGTGAACGTGGTGATCCGCG-3’ | N/A | Insig-2sg-F |
| 5’-aacCGCGGATCACCACGTTCACAC-3’ | N/A | Insig-2sg-R |
| 5’-accAACCCCTGATTGTATCCGCA-3’ | N/A | Scramble-F |
| 5’-aacTGCGGATACAATCAGGGGTT-3’ | N/A | Scramble-R |
| Primers for quantitative real-time PCR see Table S1 | N/A | N/A |
|
| ||
| Software and algorithms | ||
|
| ||
| Image Studio™ Version 5.0 | Li-COR | N/A |