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

Fig 4.

Immunofluorescence and flow cytometry of SNA and MAL II staining depict reduced fluorescence in SLC35A1 KO and NA-treated cells compared with scramble controls. Mean fluorescence intensity is significantly lower in KO cells.

SIA expression in HAE-ALI cultures differentiated from SLC35A1 KO cells. (A&B) Confocal microscopy. SNA (A) and MALII (B) lectins were used to stain glycan expression in HAE-ALISLC35A1-KO cultures. NA-treated cultures served as a positive control to show the removal of sialic acids. DyLight 649-conjugated streptavidin was used to visualize the staining under a confocal microscope at ×60 (CSU-W1 SoRa). (C&D) Flow cytometry of lection-stained cells dissociated from ALI cultures. (C) Biotinylated SNA and (D) MAL II lectins were used to stain the cell surface, followed by FITC-conjugated streptavidin for detection. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated and normalized to the WT HEK293 cells. The percentages are shown with a mean and SD from three replicates. P values as indicated were determined by using the Student’s t-test.