Nunc lab tektm 2 chamber slides

SKOV3 cells were seeded in 8-well Nunc Lab-Tek^TM II chamber slides (Thermo-Scientific, UK) at a density of 2 × 10⁴ cells/well in 300 µL of DMEM supplemented with 10% FCS, 2 mM l-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin. Cells were incubated at 5% CO₂, 37 °C and grown to ~80% confluency (24 h). Cells were washed twice with PBS, infected with 2 × 10¹⁰ phages in PBS/1% BSA in a total volume of 100 µL, diluted as appropriate by titering overnight cultures on LB/IPTG/Xgal plates. Cells were incubated for 2 h at 4 °C. Cells were washed five times in cold PBS/1%BSA/0.1% Tween-20 and incubated with mouse anti-M13 antibody (Abcam, ab9225) at a 1:50 dilution in PBS/1% BSA for 1 h at 4 °C. Bound phage were detected by incubating cells with Alexa Fluor 488 goat anti-mouse (Invitrogen, A11017) at a dilution of 1:2000 in PBS/1% BSA for 45 min at room temperature. Cells were washed four times in cold PBS/1% BSA and fixed with 4% PFA for 20 min at room temperature. Cells were washed with PBS and slides mounted with Prolong Diamond with DAPI (Invitrogen).

SKOV3 cells were seeded in 8-well Nunc Lab-Tek^TM II chamber slides (Thermo-Scientific, UK) at a density of 2 × 10⁴ cells/well in 300 µL of DMEM supplemented with 10% FCS, 2 mM l-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin. Cells were incubated in 5% CO₂, at 37 °C and grown to approximately 80% confluency (24 h). Cells were washed twice with PBS and incubated with either 0.5 mM folic acid in PBS/1% BSA or complete medium in a total volume of 300 µL/well for 1 h at 4 °C. Cells were washed with PBS and incubated with 0.5 mM FITC-labelled TVRTSAEGGCGG-COOH peptide in PBS/1% BSA in a total volume of 300 µL/well for 1 h at 4 °C. PBS was used as a no-peptide control. Cells were washed four times in cold PBS/1% BSA and fixed with 4% PFA for 20 min at room temperature. Cells were washed with PBS and slides mounted with Prolong Diamond with DAPI (Invitrogen).