(A) Cell lysates from differentiated HT29-M6 cells treated with 5-fluorouracil + irinotecan (5-FU + iri.) (50 µg/mL 5-FU + 20 µg/mL iri.) for 24 hr were analysed by Western blot with an anti-MUC5AC …
Uncropped gels for Figure 1.
(A) Schematic diagram of the experimental set-up to measure mucin production after 5-fluorouracil + irinotecan (5-FU + iri.) treatment.
(A–C) Disease-free survival (DFS) according to KChIP3 levels of CRC patients (low KChIP3 levels, n = 120; high KChIP3 levels, n = 106) (A), CRC patients with high MUC5AC levels (low KChIP3 levels, n …
(A) Expression levels of secreted mucins MUC2, MUC5AC, MUC5B, MUC6, MUC7, and MUC19. Each dot represents a different patient. Patients GSM358532, GSM358535, and GSM358538 are highlighted in green, …
(A) Comet assay of HT29-M6 cell line treated with 5-FU + iri. (25 µg/mL 5-FU + 10 µg/mL irinotecan) and 20 µM benzamil, alone or in combination. The tail moment was measured after 72 hr of treatment …
(A) Cell lysates and secreted medium of differentiated HT29-M6 cells pre-treated with a vehicle or 10 µM SN-6 inhibitor for 24 hr and then exposed to 5-FU + iri. (50 µg/mL 5-FU + 20 µg/mL iri.) for …
Uncropped gels for Figure 4.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Gene (Homo sapiens) | MUC5AC | Ensembl | ENSG00000215182 | |
Gene (H. sapiens) | MUC2 | Ensembl | ENSG00000198788 | |
Gene (H. sapiens) | MUC6 | Ensembl | ENSG00000184956 | |
Gene (H. sapiens) | MUC5B | Ensembl | ENSG00000117983 | |
Gene (H. sapiens) | MUC19 | Ensembl | ENSG00000205592 | |
Gene (H. sapiens) | KCNIP3 | Ensembl | ENSG00000115041 | |
Cell line (H. sapiens) | HT29-M6 | ATCC | CVCL_G077 | Mycoplasma free |
Cell line (H. sapiens) | HT29-18N2 | ATCC | CVCL_5942 | Mycoplasma free |
Antibody | Anti-MUC5AC (mouse monoclonal) | Neomarkers,Waltham, MA | Clone 45M1 | (1:1000) |
Antibody | Anti-γH2A.X (mouse monoclonal) | Cell Signaling | #2577 | (1:1000) |
Commercial assay or kit | CometAssay Trevigen Kit | Trevigen | 250-050K | |
Chemical compound, drug | SN-6 | Sigma-Aldrich | SML1937-5MG | (5 µM) |
Chemical compound, drug | Benzamil | Sigma-Aldrich | B2417-10MG | 5 µM |
Gene | Forward primer (5′–3′) | Reverse primer (5′–3′) |
---|---|---|
MUC5AC | CTGGTGCTGAAGAGGGTCAT | CAACCCCTCCTACTGCTACG |
TBP | TGCCCGAAACGCCGAATATAATC | GTCTGGACTGTTCTTCACTCTTGG |
GAPDH | GTCATCCCTGAGCTGAACG | CTCCTTGGAGGCCATGTG |