Background
Methods
Pancreatic tissues and animals
Cell isolation and culture conditions
Immunohistochemical staining
Host species | Dilution | Source/catalog no. | ||
---|---|---|---|---|
IHC | WB | |||
Notch1 | Rabbit | 1:50 | 1:500 | Santa Cruz/ sc-6014R |
Notch2 | Rabbit | 1:500 | 1:2000 | Lifespan/ LS-B399 |
Notch3 | Rabbit | 1:50 | 1:500 | Santa Cruz/sc-5593 |
Notch4 | Rabbit | 1:50 | 1:500 | Santa Cruz/sc-5594 |
HES1 | Rabbit | 1:200 | Santa Cruz/ sc-25392 | |
Jagged1 | Rabbit | 1:50 | Santa Cruz/ sc-8303 | |
Jagged2 | Rabbit | 1:50 | Santa Cruz/ sc-5604 | |
DLL1 | Rabbit | 1:50 | Santa Cruz/ sc-9102 | |
DLL3 | goat | 1:50 | Santa Cruz/ sc-66513 | |
DLL4 | goat | 1:50 | Santa Cruz/ sc-18640 | |
α-SMA | Mouse | 1:100 | Dako/M0851 | |
α-SMA | Rabbit | 1:100 | 1:1000 | Abcam/ab5694 |
Fibronectin | Rabbit | 1:50 | 1:1000 | Proteintech/15613–1-AP |
Collagen I | Rabbit | 1:50 | 1:1000 | Proteintech/14695–1-AP |
CSL | Rabbit | 1:1000 | Cell Signaling/5313 | |
GAPDH | Rabbit | 1:10000 | Sigma/G9545 |
Immunofluorescence
Immunocytochemical staining
siRNA-mediated Notch3 knockdown in PaSCs
Cell proliferation
Cell migration assay
RT-qPCR
Primer | Forward sequence 5′-3’ | Reverse sequence 5′-3’ |
---|---|---|
Notch1 | TCGTGCTCCTGTTCTTTGTG | CTCTCCGCTTCTTCTTGCTG |
Notch2 | GCAGGAGCAGGAGGTGATAG | ATGAGAAGCCAGGAGAGCAG |
Notch3 | TGGCTATGCTGGTGACAGTT | AGGGGGACAGGAACAGAGAT |
Notch4 | AATGCCAAGGTCAGGAACAC | AGCCCTCATCACACACACAC |
α-SMA | AATGGCTCTGGGCTCTGTAA | CTCTTGCTCTGGGCTTCATC |
Fibronectin | GAAGTCGCAAGGAAACAAGC | GTAGGTGAACGGGAGGACAC |
CollagenI | TGACTGGAAGAGCGGAGAGT | GACGGCTGAGTAGGGAACAC |
HES1 | GGCGAAGGGCAAGAATAAAT | TGCTTCACAGTCATTTCCAGA |
GAPDH | GGTTGTCTCCTGCGACTTCA | TGGTCCAGGGTTTCTTACTCC |