Fig. 5: Effect of exogenous RNase4 on the Akt-mTOR pathway in ND7/23 cells.

a, b Representative pAkt and Akt blot for each inhibitor and RNase4 (R4) condition used in culture with bar plot of mean ± SEM showing pAkt/Akt ratio quantified by western blot with n = 3 independent biological replicates. Between DMSO and R4 p = 0.00727, whereas between DMSO and all other conditions, denoted as significant p = 0.00119. For Rapamycin 5 nM and Rapamycin 5 nM +R4, p = 0.0109 and for Rapamycin 10 nM and Rapamycin 10 nM +R4, p = 0.0176. c, d Representative pmTOR and mTOR blot for each inhibitor and RNase4 condition used in culture with bar plot of mean ± SEM showing pmTOR/mTOR ratio quantified by western blot with n = 3 independent biological replicates. Between DMSO and LY294002, as well as LY294002 + R4 conditions, p = 0.00823. e, f Representative βIII tubulin blot that was run from the same gel with each phospho/total protein pair and used as loading control to normalize densitometry results prior to ratio calculations for each of the n = 3 biological replicates. Thin black lines indicate the splicing of originally non-adjacent lanes from the same blot. Inhibitors were added for 1 hour at 37 °C and include MK-2206 (Mk), LY294002 (Ly), Wortmannin (Wort), and Rapamycin (Rapa). DMSO was included as a vehicle, and 1 mg/mL RNase4 (R4) was added for 5 min after inhibitor treatment for the indicated conditions. Statistical significance between groups was determined using a two-tailed independent t-test, and the multiple hypothesis testing was corrected with Benjamini-Hochberg. Statistical significance indicated above each bar was obtained from comparisons to the DMSO group, and comparisons between RNase4 and inhibitor+RNase4 groups, as well as inhibitor and inhibitor+R4 groups, were also considered. (*p < 0.05, **p < 0.01). Source data are provided as a Source Data file.