Raw images of glycosyltransferases in Expi293F cells acquired by super-resolution confocal live imaging microscopy (SCLIM)

The folder contains separate confocal xy-plane images acquired by super-resolution confocal live imaging microscopy (SCLIM) prior to 3D reconstruction. The voxel size is 0.06 and 0.2 μm for xy and z, respectively. Colocalization of two glycosyltransferases, each fused with distinct fluorescent proteins, was analyzed. Glycosyltransferases fused with C-terminal mNeonGreen or mScarlet-I are denoted as G or R, respectively. Chimeric glycosyltransferase mutants (designated as X-Y), consisting of the N-terminal CTS region of one enzyme (X) and the C-terminal catalytic domain of another enzyme (Y), are also indicated. The SCLIM images focus on the Golgi ribbon area of Expi293F cells expressing human glycosyltransferases involved in the diversification of nonreducing terminal structures of N-glycans. This includes enzymes responsible for galactosylation [β-1,4-galactosyltransferase/B4GALT1 (GB41)], fucosylation [4-galactosyl-N-acetylglucosaminide 3-α-L-fucosyltransferase 9/FUT9], and sialylation [CMP-N-acetylneuraminate-β-1,4-galactoside α-2,3-sialyltransferase/ST3GAL3 (SA33), CMP-N-acetylneuraminate-β-galactosamide-α-2,3-sialyltransferase 4/ST3GAL4 (SA34), and β-galactoside α-2,6-sialyltransferase 1/ST6GAL1 (SA61)]. The glycosyltransferase β-1,3-galactosyltransferase/B3GALT6 (GB36), involved in GAG elongation, serves as a reference enzyme.

File Naming Convention:

• T00001: Common to all images
• C01: Raw green fluorescence image
• C02: Raw red fluorescence image
• C03: Deconvolved green fluorescence image
• C04: Deconvolved red fluorescence image
• Z001~Z065: Z-stack number (typically, the maximum Z-stack ranges between Z045 and Z065).

Within the SCLIM1 imaging data, the following subfolders are organized by individual figure folders:

• Figure 1 (E, F)

• GB36(R) vs FUT9(G)
• GB36(R) vs GB36(G)
• GB36(R) vs GB41(G)
• GB36(R) vs SA33(G)
• GB36(R) vs SA34(G)
• GB36(R) vs SA61(G)

• Figure 2 (A, B)

• GB41(R) vs GB41(G)
• GB41(R) vs SA33(G)
• GB41(R) vs SA34(G)
• GB41(R) vs SA61(G)

• Figure 2 (C, D)

• FUT9(R) vs FUT9(G)
• FUT9(R) vs GB41(G)
• FUT9(R) vs SA33(G)
• FUT9(R) vs SA34(G)
• FUT9(R) vs SA61(G)

• Figure 2 (E, F)

・SA33 Group

• SA33(R) vs SA33(G)
• SA33(R) vs SA34(G)
• SA33(R) vs SA61(G)

・SA34 Group

• SA34(R) vs SA33(G)
• SA34(R) vs SA34(G)
• SA34(R) vs SA61(G)

・SA61 Group

• SA61(R) vs SA33(G)
• SA61(R) vs SA34(G)
• SA61(R) vs SA61(G)

• Figure 4 (B, C)

• GB36(R) vs GB36(G)
• GB36(R) vs GB61-GB41(G)
• GB36(R) vs GB41-GB36(G)

• Figure 4 (D, E)

• SA61(G) vs SA34-SA61(R)
• SA61(G) vs SA61(R)

• Figure 4 (F, G)

• SA34(G) vs SA34(R)
• SA34(G) vs SA34-SA61(R)

• Figure 4 (H, I)

• SA34(G) vs SA61-SA34(R)
• SA61(G) vs SA61-SA34(R)