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. 2022 Oct 31;33:100926. doi: 10.1016/j.ymgmr.2022.100926

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Fig. 1 — Expression of α-Gal A isoforms in transfected HEK293 GLA^−/−cells. (A) Representative Western blot showing the absence of α-Gal A in Crispr/Cas9 edited cells. GAPDH was used as loading control. (B) Schematic representation of human α-Gal A. The grey box represents the leader sequence; the light blue trident represent glycosylation sites, represent active sites, the yellow box represents the substrate binding site. The position of selected variants is shown. In yellow, variants with residual activity; in red, variants with no residual activity. The residual activity of D231N variant is reported 0 or 0.5% of wild type activity [5,9]. (C) α-Gal A expression in transiently transfected HEK293 GLA^−/−cells assessed by real-time RT-qPCR. Expression is normalized to HPRT1. Data are expressed as mean ± s.d. (n = 3 independent experiments). (D) Western blot analysis showing α-Gal A expression in lysates of HEK293 GLA^−/− cells transfected with different α-Gal A isoforms. Actin is shown as a loading control.

Expression of α-Gal A isoforms in transfected HEK293 GLA^−/−cells. (A) Representative Western blot showing the absence of α-Gal A in Crispr/Cas9 edited cells. GAPDH was used as loading control. (B) Schematic representation of human α-Gal A. The grey box represents the leader sequence; the light blue trident represent glycosylation sites, represent active sites, the yellow box represents the substrate binding site. The position of selected variants is shown. In yellow, variants with residual activity; in red, variants with no residual activity. The residual activity of D231N variant is reported 0 or 0.5% of wild type activity [5,9]. (C) α-Gal A expression in transiently transfected HEK293 GLA^−/−cells assessed by real-time RT-qPCR. Expression is normalized to HPRT1. Data are expressed as mean ± s.d. (n = 3 independent experiments). (D) Western blot analysis showing α-Gal A expression in lysates of HEK293 GLA^−/− cells transfected with different α-Gal A isoforms. Actin is shown as a loading control.