Fig. 3: Downregulation of PC core proteins impairs autophagy machinery in hippocampal neurons.
From: A primary cilia–autophagy axis in hippocampal neurons is essential to maintain cognitive resilience
a, LC3B and SQSTM1/p62 immunofluorescence and puncta quantification performed on brain cross-sections at the level of the hippocampal CA3 region from 3-month-old mice, 3 weeks after local stereotactic injections with AAV-U6-shRNA-Ift20, AAV-U6-shRNA-Kif3a or AAV-U6-shRNA-scramble. Scale bar, 20 µm. b, Representative western blot image and quantification of LC3B-II accumulation in 3-month-old mouse hippocampi, 3 weeks after local stereotactic injections with either eSYN-shRNA-Ift20mir or eSYN-shRNA-scramble-mir (two independent cohorts). β-actin was used as a loading control for each sample. c, SQSTM1/p62 immunofluorescence (blue) and punctae quantification in the CA3 region of the hippocampi of 3-month-old mice, 3 weeks after local stereotactic injections of AAVs expressing either eSYN-shRNA-Ift20-mir or eSYN-shRNA-scramble-mir. NeuN staining (purple) was used to label neuronal nucleus. Scale bar, 25 µm. Data were obtained from two independent cohorts. d, PC immunofluorescent staining (green, stained by ACIII) and PC length measurement in the CA3 region of mouse after stereotactic injections with either eSYN-shRNA-Ift20-mir or shRNA-scramble-mir. The length of the PC was measured and quantified in 3D using Imaris software. The length is expressed in µm, and 2–4 brain sections (n = 50–150 neurons per section) were analyzed per mouse. Scale bar, 10 µm. All data are expressed as mean ± s.e.m., and P values were determined by one-tailed (a) or two-tailed (b–d) Student’s t-tests compared to Scrb.
