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. 2023 Feb 2;51(4):1583–1599. doi: 10.1093/nar/gkad023

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Simultaneous PIKfyve and ARF6 inhibition is both necessary and sufficient to account for the increase in ASO and siRNA activity in SH-BC-893-treated cells. (A) MALAT1 levels in HeLa cells treated with the indicated concentrations of cEt gapmer ASO targeting MALAT1 ± SH-BC-893 (5 μM), NAV2729 (12.5 μM), YM201636 (800 nM), or both for 24 h. Mean ± SD shown, n = 3. (B) IC₅₀s from each biological replicate in (A); mean ± SD shown. Due to unequal SD, a Brown–Forsythe and Welch ANOVA test was used with Dunnett's T3 test to correct for multiple comparisons; **P < 0.01. (C) HPRT1 mRNA levels in HeLa cells treated with the indicated concentrations of a palmitate-conjugated siRNA targeting HPRT1 ± SH-BC-893 (5 μM), NAV2729 (12.5 μM), apilimod (100 nM) or NAV2729 and apilimod for 24 h. Mean ± SD shown, n = 3. (D) IC₅₀s from each biological replicate in (C); mean ± SD shown. Using an ordinary one-way ANOVA with Sidak's multiple comparison test, **P < 0.01; ***P < 0.001. (E) MALAT1 levels in HeLa cells stably expressing luciferase or GRP1^DD treated with the indicated concentrations of cEt ASO targeting MALAT1 ± SH-BC-893 (5 μM) for 24 h. Mean ± SD shown, n = 3. (F) IC₅₀s from each biological replicate in (E); mean ± SD shown. (G) MALAT1 levels in HeLa cells treated with the cEt ASO targeting MALAT1 ± SH-BC-893 (5 μM) or PPZ (15 μM) for 24 h. Mean ± SD shown, n = 3. (H) IC50s from each biological replicate in (G); mean ± SD shown. Because SD are not equal, Brown-Forsythe and Welch ANOVA test was used with Dunnett's T3 test to correct for multiple comparisons; **P < 0.01.