Skip to main content
. 2019 May 2;5(3):e337. doi: 10.1212/NXG.0000000000000337

Figure 5. Expression and localization of HA-tagged TNPO3 constructs.

Figure 5

(A) Schematic of HA-tagged TNPO3-WT, the previously reported LGMD1F mutation TNPO3+15 or the currently described frameshift mutation TNPO3-RG. (B) Immunoblot with anti-HA, TNPO3, or GAPDH of HeLa cell lysates transfected with HA-TNPO3-WT, HA-TNPO3+15, or HA-TNPO3-RG. (C) HA immunofluorescence of HeLa cells transiently transfected with HA-TNPO3-WT, HA-TNPO3+15, or HA-TNPO3-RG. HA-positive cytosolic puncta (arrows). (D) Quantitation of the percent of cells in C with cytosolic HA-TNPO3 puncta (*** p = < 0.0001). (E) Immunofluorescence using antibodies to HA (green) and TIA1 (red) of HeLa cells transfected with TNPO3-WT or TNPO3-RG pre- and post-stress granule induction with arsenite. HA-positive/TIA1-negative puncta (arrowheads). (F) Immunofluorescence using antibodies to HA (red) and RanGAP1 (green) or nucleoporin antibody Mab414 (green) of HeLa cells transfected with TNPO3-WT or TNPO3-RG. Cytosolic puncta (arrowheads). Scale bars (C, E, and F) = 20 μm. TNPO3 = transportin-3; WT = wild type.