Fluorescent microscopy was performed as previously described21 (link). H9C2 cells split onto sterile cover glasses coated with poly-L-lysine (Sigma) were transfected with adenoviruses encoding wildtype or lysine mutant XBP1s-GFP fusion proteins and then incubated with HG for 6 hours. Cells cover glasses were removed from dishes and then counterstained with DAPI. The fluorescent images were taken with a Nikon Eclipse E600 microscope or a Zeiss Axiovert 135.
Immunofluorescence microscopy was performed as previously described27 (link). Briefly, H9C2 cells on coverslips were fixed with 4% paraformaldehyde for 15 min, then washed with PBS three times. The cells were blocked with normal goat serum and incubated with anti-XBP1s antibody overnight. Cells were washed with PBS three times and then incubated with secondary antibodies conjugated with Alexa Fluor 488 goat anti-mouse IgG antibody (Invitrogen) for 1 hour. Cells were washed with PBS three times and then counterstained with DAPI. Immunofluorescence images were taken with a Nikon Eclipse E600 microscope or a Zeiss Axiovert 135.
Immunofluorescence microscopy was performed as previously described27 (link). Briefly, H9C2 cells on coverslips were fixed with 4% paraformaldehyde for 15 min, then washed with PBS three times. The cells were blocked with normal goat serum and incubated with anti-XBP1s antibody overnight. Cells were washed with PBS three times and then incubated with secondary antibodies conjugated with Alexa Fluor 488 goat anti-mouse IgG antibody (Invitrogen) for 1 hour. Cells were washed with PBS three times and then counterstained with DAPI. Immunofluorescence images were taken with a Nikon Eclipse E600 microscope or a Zeiss Axiovert 135.
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