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. 2024 Nov 27;16(1):e03268-24. doi: 10.1128/mbio.03268-24

Fig 7.

Immunofluorescence images of HEK293 and HAE-ALI cells depicting TGN46, SLC35A1, and rAAV capsid staining. SLC35A1 KO cells exhibit reduced SLC35A1 and altered capsid localization compared to scramble controls, highlighting localization differences.

SLC35A1 and AAV capsid are colocalized with TGN46. (A and B) HEK293 cells. SLC35A1-KO or Scramble HEK293 cells were transduced with (A) rAAV5 at MOI of 20,000 or (B) rAAV2.5T at MOI of 2,000. At 8 hpt, the cells were fixed and permeabilized, followed by immunostaining with the first antibody against indicated protein and fluorescence-conjugated secondary antibodies. (C) HAE-ALI cultures. The HAE-ALI cultures differentiated from SLC35A1-KO or Scramble CuFi-8 cells were transduced with rAAV2.5T at a MOI of 20,000. At 3 dpt, the cells were fixed and permeabilized, followed by immunostaining with the first antibody against indicated protein and fluorescence-conjugated secondary antibodies. The stained cells were imaged under a confocal microscope (CSU-W1 SoRa, Nikon) at 60× with 4× SoRa magnitude (scale bar = 20 µm).