Action of compound 25o as pharmacological chaperone
for common pathogenic AGT variants. CHO-GO-AGTwt, CHO-GO-AGT-Mi, CHO-GO-G170R-Mi,
CHO-GO-I244T-Mi, and CHO-GO-F152I-Mi cells were grown for 7 days in
the absence or presence of 50 μM 25o and 10 μM
PN. At the end of treatment, cells were detached and lysed, and the
soluble fraction of each sample used for (A) AGT enzymatic activity
determination. The AGT specific activity of control cells of each
clone was assumed to be 1 to help assess the changes (CHO-GO-AGTwt
cells: 191 ± 10 nmol of pyruvate/min/mg protein; CHO-GO-AGT-Mi:
132 ± 9 nmol of pyruvate/min/mg protein; CHO-GO-G170R-Mi: 54
± 7 nmol of pyruvate/min/mg protein; CHO-GO-I244T-Mi: 28 ±
5 nmol of pyruvate/min/mg protein; CHO-GO-F152I-Mi 24 ± 5 nmol
of pyruvate/min/mg protein). Data represent mean ± SEM (n = 3). *p < 0.05 vs respective control
cells. #p < 0.05 vs respective PN-treated
cells. (B) AGT expression level quantification. AGT levels in control
cells of each clone were assumed to be 1 to help assess the changes.
GAPDH has been used as loading control. The images are representative
of one out of three separate experiments. Data represent mean ±
SEM (n = 3). *p < 0.05 vs respective
control cells. #p < 0.05 vs respective
PN-treated cells.