EGFP positive cells are disorganized and have increased EGFP signals in mGFAP:CKO cerebellums.

<p>(a) In symptomatic 2 month old mGFAP:CKO mice, massive cell loss was observed in mutant cerebellum (arrows indicate missing cells in cerebellar lobes). (b-d) EGFP positive cells had up-regulated signals and appeared in the molecular layers in the mutant cerebellums. (e) qRT-PCR also indicated up-regulated levels of <i>Aldh1l1</i> transcripts (p-value< 0.01, n = 3). (e) <i>Gfap</i> showed an upward trend suggesting astrogliosis (p-value = 0.05, n = 3). (e) <i>Aqp4</i> levels were similar between wildtype and mutant animals (p-value = 0.14, n = 3). (e) <i>Glast</i> level was decreased in mutant cerebellums (p-value < 0.01, n = 3). (f-h) In P16 mGFAP:CKO, defects with Aldh1l1-EGFP could already be observed in the cerebellum. Aldh1l1-EGFP cells were present in mutant molecular layer. (i-k) While ki67 positive cells were limited in wildtype cerebellum, we saw a noticeable increase in mutant cerebellum (ki67 cells are red). (l-m) These ki67 positive cells co-localized predominately with EGFP cells. <i>Aldh1l1</i> and <i>Gfap</i> transcripts were up-regulated while <i>Glast</i> had a small decrease expression level in mutant cerebellum. (o) No change was detected in <i>Aqp4</i> (<i>Aldh1l1</i> p-value< 0.01, <i>Aqp4</i> p-value = 0.16, <i>Gfap</i> p-value< 0.05, and <i>Glast</i> p-value< 0.05, n = 3). Scale bars in a-d and f-n represent 50 μm. Gray bars represent wildtype mice and yellow bars represent mGFAP:CKO mice in e and o. Asterisks in e and o indicate significant level (** represents p< 0.01 and * represents p< 0.05). Y axes in e and o represent the levels for gene of interest normalized to <i>Gapdh</i>. The average expression level of wildtype samples were normalized to 1. Unpaired t-test was used to determine p-values.