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. Author manuscript; available in PMC: 2022 May 23.
Published in final edited form as: Biochim Biophys Acta Mol Basis Dis. 2020 Jun 25;1866(10):165883. doi: 10.1016/j.bbadis.2020.165883

Figure 3. CLN3 deficiency led to autophagy-inducing mTORC1/Akt suppression and AMPK activation in RPE-1 cells.

Figure 3.

(A) Representative immunoblotting data showed that CLN3 siRNA treatment of RPE-1 cells resulted in decreased S6K phosphorylation at T389, decreased Akt phosphorylation at T308 and S473, and increased AMPK phosphorylation at T172. Total S6K, Akt and AMPK protein levels as well as actin levels were largely unchanged. (B) Quantification of immunoblotting results showed decreased phospho-S6K (T389)/total S6K ratio that suggests mTORC1 suppression in response to CLN3 exon 8 siRNA (versus non-targeting siRNA) treatments under both non-starved (open bars) and 18 h FBS-starved (closed bars) conditions. (C) Quantification of immunoblotting results showed increased phospho-AMPK (T172)/total AMPK ratio that suggests AMPK activation in response to CLN3 exon 8 siRNA (versus non-targeting siRNA) treatments under both non-starved (open bars) and 18 h FBS-starved (closed bars) conditions. Quantification of immunoblotting results showed decreased (D) phospho-Akt (T308)/total Akt and (E) phospho-Akt (S473)/total Akt ratios that suggest Akt suppression in response to CLN3 exon 8 siRNA (versus non-targeting siRNA) treatments under non-starved conditions. Four-six independent experiments were averaged. Statistics were carried out using t-test.