Fig. 4: GALNTL5 localization to the ER impairs calnexin function. | Cell Death Discovery

Fig. 4: GALNTL5 localization to the ER impairs calnexin function.

From: GALNTL5, which is restricted to mouse spermatids, impairs endoplasmic reticulum (ER) function through direct interaction with ER chaperone proteins

Fig. 4

A, B Fluorescent images of GALNTL5-GFP/GC-2spd cells stably expressing NHK tagged with DsRed2. Without the induction of GALNTL5-GFP expression, the NHK-DsRed signals co-localize with calnexin in the ER (A). When the expression of GALNTL5-GFP was induced with a cumate solution, NHK-DsRed signals disappeared from the ER 2 days later but those of GALNTL5-GFP persisted (B). Nuclei are stained with DAPI. White scale bars in the merged image, 20 µm. C Transient expression of NHK tagged with Flag in GALNTL5-GFP/GC-2spd cells. NHK-Flag signals decreased with persistent, 2-day GALNTL5 expression in GALNTL5-GFP/GC-2spd cells. Histone-H3 was used as the control. D Quantification of band intensity of NHK-Flag signals were statistically analyzed (mean ± SEM. n = 3. Two-tailed Student’s t-test. **p < 0.01). E Western blot of calnexin and NHK-DsRed (NHK protein tagged with DsRed) in GC-2spd(ts) cells expressing NHK-DsRed at 72 h after calnexin siRNA induction. Histone-H3 was used as the control. F The alterations of protein levels of calnexin and NHK-DsRed were quantified (mean ± SEM. n = 3. Two-tailed Student’s t-test. **p < 0.01, ***p < 0.001).

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