Fig. 3
GABRA1 and NPTN interaction in the primary neurons and hippocampus tissues of rats and GABRA1 endocytosis a Immunofluorescence co-localization analysis of primary neurons (microscopes, Olympus BX53; objectives, neurons; time of exposure, 50 ms; optic/digital magnification conditions, × 600): GABRA1 co-localized with NPTN in neurons (PCCCtrl = 0.83, PCCMg2+-free = 0.91). Scale bar: 50 μm. b Immunofluorescence co-localization analysis of hippocampus tissue (microscopes, Olympus BX53; objectives, hippocampal tissues; time of exposure, 250 ms; optic/digital magnification conditions, × 200): GABRA1 co-localized with NPTN in hippocampus tissue (PCC Ctrl = 0.61, PCCSE = 0.73). Scale bar: 500 μm. c Co-IP (in vitro): Pooled quantification of protein immunoprecipitation shows a significant reduction in the pull-down of NPTN in the Mg2 + -free group compared with the control group (n = 6 in each group, #p < 0.01) and significant increase in the pull-down of NPTN in the LV-OE-NPTN (Mg2 + -free) group compared with the LV-NC (Mg2 + -free) group (n = 5 in each group, #p < 0.05). d Co-IP (in vivo): Pooled quantification of protein immunoprecipitation shows a significant reduction in the pull-down of the NPTN in the SE group compared with the control group (n = 6 in each group, *p < 0.05) and a significant increase in the pull-down of NPTN in the AAV-OE-NPTN (SE) group compared with the AAV-NC (SE) group (n = 5 in each group, *p < 0.05). e The intracellular accumulation ratio of neurons in the LV-OE-NPTN (Mg2 + -free) group was decreased significantly compared with that of those in the control group (intracellular accumulation ratio LV-NC (Mg2+-free) = 0.76 ± 0.05, intracellular accumulation ratio LV-NC (Mg2+-free) = 0.47 ± 0.06, n = 9 in each group, #p < 0.01). Scale bar: 50 μm. All experiments were repeated for 3 times. △Independent t-tests were used for the comparison of means