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. 2021 May 26;11:10962. doi: 10.1038/s41598-021-90554-3

Figure 2.

Figure 2

Analysis of cell type-specific markers by qRT-PCR in embryonic mouse retinal explants treated with neuropeptide agonists and antagonists. Cell type marker genes tested include Ccnd1 (all progenitors); Fgf15 and Vsx2 (primary progenitors); Neurog2 and Atoh7 (neurogenic progenitors); Otx2 and Crx (photoreceptor precursors); Pou4f2 (retinal ganglion cells); Penk, Sstr2, and Gal (neuropeptides in the study, with each of these also being selective markers of neurogenic progenitors); and Gapdh (internal control). Each point represents a sample (average of 3 technical replicates). Each sample was tested for each gene. (A) Marker gene expression relative to average control with Npy treatment. Concentration = 1 μM. n = 3 each condition. Explants treated E18-P2. (B) Marker gene expression relative to average control with Galanin or M40 treatment. Concentration = 1 μM. n = 6 each condition. Explants treated E14-P0. (C) Marker gene expression relative to average control with Met-Enkephalin or Naltrexone treatment. Concentration = 500 nM. n = 3 each condition. Explants treated E14-P0. (D) Marker gene expression relative to average control with treatment with SSTR2 agonist (1R,1'S,3'R/1R,1'R,3'S)-L-054,264. Concentration: 500 nM. n = 3 each condition. Explants treated E14-P0. (E) Otx2 expression relative to average control of explants treated with increasing levels of Sstr2 agonist. The trend line represents a linear model, with gray area denoting the 95% confidence interval. n = 6 each condition (except for the 5 nM treatment, where n = 4). Explants treated between E14-P0. (F) Crx expression relative to average control of explants treated with increasing concentrations of Sstr2 agonist. Trend line represents a linear model with the gray area denoting 95% confidence interval. n = 6 each condition (except for 5 nM treatment, where n = 4). Explants treated from E14-P0. (G) Otx2 expression relative to average control of Sstr2−/− explants treated with 500 nM Sstr2 agonist. n = 6 each condition. (H) Crx expression relative to controls in Sstr2−/− explants treated with 500 nM Sstr2 agonists. n = 6 each condition. Explants treated E14-P0. (A–D) P-value calculated using ANOVA. + indicates a nominally significant P-value below 0.05. *Indicates a P-value significant at a Bonferroni correction below 0.0045. (E, F) P-value calculated with linear model of expression ~ condition + RNA extraction batch. (G, H) P-value calculated using student’s t-test.