REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rat-anti-IGFBP2 | R&D systems | Cat# MAB7971; RRID:AB_2264598 |
Rabbit anti-IGFBP2 for IHC | Abcam | Cat# ab188200 |
Rabbit anti-P21 | Abcam | Cat# ab188224; RRID:AB_2734729 |
Rabbit anti-Collagen 1 | EMD Millipore | Cat# AB765P; RRID:AB_92259 |
Rabbit anti-PPARα | LSBio | Cat# LS-C312574 |
Rabbit anti-Fibronectin | Abcam | Cat# ab2413; RRID:AB_2262874 |
Rabbit anti-Vimentin | CST | Cat# 5741S; RRID:AB_10695459 |
HRP conjugated-β actin | Santa Cruz | Cat# sc-47778HRP; RRID:AB_2714189 |
Mouse anti-α tubulin | CST | Cat# 3873; RRID:AB_1904178 |
Rabbit anti-H3 Histone | CST | Cat# 4499; RRID:AB_10544537 |
Rabbit anti-phospho-Histone 2AX | CST | Cat# 2577; RRID:AB_2118010 |
Rabbit anti-Prosurfactant Protein C | Abcam | Cat# ab211326; RRID:AB_2927746 |
Bacterial and virus strains | ||
Ad-Null | Vector Biolabs | Cat#1240 |
Ad-m-TGFβ1 | Vector Biolabs | Cat#ADV-274099 |
mouse Igfbp2 lentiviral particle | Origene | Cat#: MR204287L3V |
Lenticontrol particles | Origene | Cat# PS100092V |
Chemicals, peptides, and recombinant proteins | ||
human IGFBP2 recombinant protein | R&D Systems | Cat# 797-B2-025 |
Bleomycin | EMD Millipore | Cat# 203401 |
DMSO | Sigma- Aldrich | Cat# D8418 |
4% PFA | Santa Cruz | Cat# sc-281692 |
DMEM/F12 | Gibco | Cat# 11-320-033 |
Plasmocin | invivogen | Cat# ant-mpp |
Collagenase type 1 | Worthington Biochemical | Cat# LS004197 |
Dispase | Corning | Cat# 354235 |
Green chromogen | Leica | Cat# DC9913 |
Bond polymer refine system | Leica | Cat# DS9800 |
BondTM Primary antibody diluent | Leica | Cat# AR9352 |
BOND Epitope Retrieval Solution 1 | Leica | Cat# AR9961 |
BOND Epitope Retrieval Solution 2 | Leica | Cat# AR9640 |
Wash solution 10X concentration | Leica | Cat# AR9590 |
BOND Universal Covertile | Leica | Cat# S21.4611 |
Tween-20 | Sigma- Aldrich | Cat# P9416 |
PVDF membrane | EMD Millipore | Cat# IPVH00010 |
SuperSignalTM Pico or Femto substrate | Thermo Fisher Scientific | Cat# 34095 |
RIPA buffer | Thermo Fisher scientific | Cat# 89901 |
Igfbp2 | Life science technology | Hs01040719_m1 |
18S rRNA | Life science technology | Hs99999901_s1 |
Pparα | Life science technology | Mm00440939_m1 and Hs00231882_m1 |
Igfbp2 siRNA | Horizon Discovery | L-062198-01-0005 |
Pparα siRNA | Santa Cruz | sc-36308 |
Critical commercial assays | ||
High-Capacity RNA-to-cDNA™ Kit | Thermo Fisher scientific | Cat# 4387406 |
TaqMan™ Fast Advanced Master Mix | Thermo Fisher scientific | Cat# 44-445-56 |
Hydroxyproline assay | Cell Biolab | Cat# STA-675 |
Βeta-galactosidase activity assay | Biovision | Cat# K821- 100 |
RT2 profiler PCR assay | Qiagen | Cat# PAMM-050ZC-6 |
IGFBP2 ELSA assay | R&D systems | Cat# DY797 |
BCA protein assay | Thermo Fisher Scientific | Cat# 23227 |
Experimental models: Cell lines | ||
MLE-12 | ATCC | CRL-2110 |
Experimental models: Organisms/strains | ||
Igfbp2flx/flxtransgenic mouse | Leeds University | |
Sftpc-CreERT2 | The Jackson Laboratory | RRID#IMSR_JAX:028054 |
C57BL6/J | The Jackson Laboratory | RRID#IMSR_JAX:000664 |
Oligonucleotides | ||
IL-1βCCH | PrimerBank ID 6680415a1 | 5′-GCAACTGTTCCTGAACTCAACT-3′ and 5′-ATCTTTTGGGGTCCGTCAACT-3′ |
IL-4 | PrimerBank ID 10946584a1 | 5′- GGTCTCAACCCCCAGCTAGT-3′ 5′- GCCGATGATCTCTCTCAAGTGAT-3′ |
IL-6 | PrimerBank ID 13624311a1 | 5′-TAGTCCTTCC-3′ 5′-TACCCCAATTTCC-3′ |
MCP-1 | PrimerBank ID 6755430a1 | 5′- TTAAAAACCTGGATCGGAACCAA-3′ 5′- GCATTAGCTTCAGATTTACGGGT-3′ |
STAT-3 | PrimerBank ID 13277852a1 | 5′- CAATACCATTGACCTGCCGAT-3′ 5′- GAGCGACTCAAACTGCCCT-3′ |
STAT6 | PrimerBank ID 6678155a1 | 5′- CTCTGTGGGGCCTAATTTCCA-3′ 5′- CATCTGAACCGACCAGGAACT-3′ |
TNF-α | PrimerBank ID 7305585a1 | 5′- CCCTCACACTCAGATCATCTTCT-3′ 5′- GCTACGACGTGGGCTACAG-3′ |
Software and algorithms | ||
ImageJ 1.53t | FIJI | https://imagej.nih.gov/ij/ |
Graphpad Prism 9.0 | Graphpad software LLC | https://www.graphpad.com |