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Fig. 3 | Molecular Brain

Fig. 3

From: Cirbp-PSD95 axis protects against hypobaric hypoxia-induced aberrant morphology of hippocampal dendritic spines and cognitive deficits

Fig. 3

Hypoxic exposure resulted in aberrant dendritic spine morphology and PSD95 expression in vivo and Cirbp regulated PSD95 expression through 3′UTR binding. A Left: representative images of hippocampal neurons with hypoxia or normoxia exposure, scale bar = 10 μm. Middle: enlarged images of hippocampal neurons, red triangle indicated the elimination of spines or filopodia, scale bar = 5 μm. Right, the quantification of spines density (n = 6 per group, Student’s t-test, ± SEM). B Morphological changes of primary hippocampal neurons under hypoxic exposure, scale bar = 50 μm. C Representative immunoblotting (left) and quantitative analysis (right) of PSD95 in primary hippocampal neurons under hypoxic exposure (n = 3 biological replicates, Student’s t-test, ± SEM). D The relative mRNA levels of PSD95 in primary hippocampal neurons under hypoxic exposure (n = 3 biological replicates, Student’s t-test, ± SEM). E Representative immunoblotting and quantitative analysis of PSD95 in HT-22 cells following normoxic or hypoxic exposure (n = 3 biological replicates, Student’s t-test, ± SEM). F The relative mRNA levels of PSD95 in HT-22 cells (n = 3 biological replicates, Student’s t-test, ± SEM). G PSD95 expression after CHX treatment based on Western blot and quantitative analysis (n = 3 biological replicates, paired Student’s t-test, ± SEM). H RBP binding motif distributing in PSD95 3′UTR and construction of a series of luciferase reporter vectors containing PSD95 3′UTR fragments. I Luciferase activity assays to examine the functional RBP motif sites in the PSD95 3′UTR regulated by Cirbp (n = 3 biological replicates, Student’s t-test, ± SEM). JL The protein levels of Cirbp in mice hippocampus (J), primary hippocampal neurons (K) and HT-22 cells (L) following hypoxia exposure (n = 3 biological replicates, Student’s t-test, ± SEM). M RNA immunoprecipitation (RIP) followed by RT-PCR in HT-22 cells (n = 4 biological replicates, Student’s t-test, ± SEM). *p < 0.05, **p < 0.01, ***p < 0.001

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