Lab tek 4 well chamber slides

JFH1/GFP-transfected Huh7 cells grown on Lab-Tek 4-well chamber slides (Nunc, Roskilde, Denmark) were fixed with 4% paraformaldehyde (Sigma-Aldrich) for 15 min. After washing three times with PBS, nuclei were visualized by staining with 1 μM 4′,6′-diamidino-2-phenylindole (DAPI). Fluorescent signal was visualized using a confocal laser scanning microscope (Zeiss LSM 510 META, Carl Zeiss, Oberkochen, Germany). The GFP-positive area among 500 total cells was quantified using ImageJ program (https://imagej.nih.gov/ij/).

22Rv1 cells were plated on Lab-Tek 4-well chamber slides (Nunc, Naperville, IL) and cultured to approximately 80% confluency. Cells were treated with compounds for 24 hours, fixed with 3.7% formaldehyde, washed with PBS, permeabilized for 10 minutes with 0.1% Triton-X 100, and blocked with 5% BSA for 30 minutes. Slides were then incubated overnight at 4 °C with primary antibodies, followed by incubation with fluorescent secondary antibodies for 2 hours at room temperature in a dark, humidified chamber. Cover slips were mounted on slides with ProLong Gold Antifade Mountant containing DAPI (Life Technologies). Fluorescent images were acquired using a Zeiss LSM 710 confocal laser-scanning microscope using 40x or 63x objectives. Acquired images were enhanced for brightness and contrast.