SKOV-3 cells
(HTB-77, passage
81) and A549 cells (CCL-185, passage 27) were purchased from ATCC
(American Type Culture Collection, Manassas, VA 20108 USA). HAL-15,
a primary cell line isolated from normal human liver, was a kind gift
from Prof. George Yeoh, the University of Western Australia. All cells
were maintained in folate free RPMI 1640 cell culture medium (GIBCO)
supplemented with 100 units/mL penicillin G, 100 μg/mL streptomycin
(Sigma Aldrich), and 10% fetal bovine serum (Invitrogen). For immunofluorescence,
the primary antibody used was an α-tubulin (DM1A) mouse mAb
(Cell signalling technologies) and the secondary antibody used was
an AlexaFlour 488 Goat Anti-Mouse IgG antibody (Molecular Probes).
DAPI was used for nuclei staining. Antibodies used for the flow cytometry
analysis of receptor expression were allophycocyanin (APC) human FOLR1
mAb (FRα) antibody (R&D systems) and APC-tagged mouse IgG1
K Isotype control (R&D systems). The antibody diluent used was
phosphate-buffered saline (PBS) with 10% normal goat serum (Invitrogen)
and 0.1% Triton-X (Sigma Aldrich).
(HTB-77, passage
81) and A549 cells (CCL-185, passage 27) were purchased from ATCC
(American Type Culture Collection, Manassas, VA 20108 USA). HAL-15,
a primary cell line isolated from normal human liver, was a kind gift
from Prof. George Yeoh, the University of Western Australia. All cells
were maintained in folate free RPMI 1640 cell culture medium (GIBCO)
supplemented with 100 units/mL penicillin G, 100 μg/mL streptomycin
(Sigma Aldrich), and 10% fetal bovine serum (Invitrogen). For immunofluorescence,
the primary antibody used was an α-tubulin (DM1A) mouse mAb
(Cell signalling technologies) and the secondary antibody used was
an AlexaFlour 488 Goat Anti-Mouse IgG antibody (Molecular Probes).
DAPI was used for nuclei staining. Antibodies used for the flow cytometry
analysis of receptor expression were allophycocyanin (APC) human FOLR1
mAb (FRα) antibody (R&D systems) and APC-tagged mouse IgG1
K Isotype control (R&D systems). The antibody diluent used was
phosphate-buffered saline (PBS) with 10% normal goat serum (Invitrogen)
and 0.1% Triton-X (Sigma Aldrich).