To assess LAMP-1 immunoreactivity in α-Gal A-deficient mouse brain we first performed chromogenic detection.
The effect of a range of inhibitors known to affect Taq polymerase [ 25, 27] was tested on LAMP-BART to assess whether they affect the Bst polymerase used in LAMP or the LAMP-BART reaction couple.
LAMP-2 knockout mice and wild-type controls (both: n = 4) were examined in a battery of behavioral tests to assess functional consequences of LAMP-2 deficiency.
To assess whether loss of LAMP-2 leads to lysosomal dysfunction that correlates with the observed neuropathological abnormalities, we determined enzyme activities of the lysosomal hydrolases β-hexosaminidase and β-glucuronidase as well as the maturation of cathepsin D as a means of evaluating lysosomal function.
The amount of DCs did not correlate with the anterior chamber cell count or anterior chamber flare, assessed by slit lamp examination (data not shown).
The quality of the lenses was assessed using slit lamp biomicroscopy and only lenses of good optical quality were used for the experiments.
Structures of the eye were assessed with slit lamp biomicroscopy examination.
Both eyes had no inflammatory signs in the anterior segment and vitreous, as assessed by slit lamp biomicroscopy examination.
An illuminator was positioned behind the examinees' ears; the angle between the illuminator and the microscope was 30 ~ 45 degrees with a 10× magnification.Without iridodilator usage, the characteristics of lens were assessed using slit lamp with proper brightness, height, and width.
The lenses were assessed by slit lamp examination after pupil dilatation and graded using the grading scale for opalescence from 0 to III for nuclear cataract, from 0 to IV for cortical cataract, and from 0 to III for posterior sub capsular cataract.
Since prolonged starvation under nutrient free conditions induces expression and nuclear translocation of the transcription factor EB (TFEB), that is known to modulate lysosomal biogenesis [ 46], we have assessed protein levels of LAMP-2A, LAMP-2B, LAMP-1 and LIMP-2 via immunoblotting and qRT-PCR analysis.
