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. 2023 Jun 28;43(26):4907–4925. doi: 10.1523/JNEUROSCI.1894-22.2023

Figure 2.

Figure 2.

Atg8a overexpression in PDF+ lateral ventral neurons does not improve the levels of PDF neuropeptide in the small LNvs (s-LNv). A, Representative MIP images (day 1, adult brain) of PDF neurons (arrow indicates large neurons; dotted circles represent small neurons) depicting staining of GFP-Atg8a (green), mutant HTT-Q128 (red), and PDF (magenta). Right top diagram represents the right half of the Drosophila brain depicting the position of PDF neurons in the brain. Scale bar, 20 µm. (+) and (–) in the bar graphs represent the presence or absence, respectively, of the gene in the fly. B, Plot represents quantification of HTT protein intensity from small and large neurons when only HTT-Q0 protein or HTT-Q0 and Atg8a protein were coexpressed. No significant decrease in the intensity of HTT protein was observed on the expression of two UAS constructs in the PDF neurons. n > 16 brain hemispheres. *p < 0.05. C, Plot represents % rhythmicity values for control and experimental genotypes. Control flies were rhythmic; however, no significant improvement in % rhythmicity was observed when mutant HTT-Q128 protein was coexpressed with UAS-GFP. n > 17 flies/genotype. *p < 0.05. D, Plot represents quantification of the number of PDF+ small neurons. Based on PDF neuropeptide and HTT-Q128 staining, quantification was done at three different ages (days 1, 5, and 10). Control genotype showed ∼4 PDF+ small neurons per hemisphere. Compared with the control genotype, mutant HTT-Q128 protein expression led to a significant reduction in the number of small neurons. A small improvement in the number of small neurons was observed on Atg8a overexpression. n > 12 brain hemispheres (for control genotype), and n > 16 brain hemispheres (for experimental genotype). *p < 0.05. E, Plot represents quantification of the number of PDF+ large neurons. Based on PDF neuropeptide and HTT-Q128 staining, quantification was done at three different ages (days 1, 5, and 10). Control genotype showed ∼4 PDF+ large neurons per hemisphere. Compared with the control genotype, mutant HTT-Q128 protein expression in the large neurons does not lead to any significant reduction in the number of neurons. Neither any change was observed on Atg8a overexpression in the presence of mutant HTT-Q128 protein. n > 12 brain hemispheres (for control genotype), and n > 16 brain hemispheres (for both experimental genotypes). *p < 0.05. Asterisk on individual genotypes indicates that the genotype is significantly different from all other plotted genotypes.