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. 2020 Sep 1;48(17):9822–9839. doi: 10.1093/nar/gkaa699

Figure 3.

Figure 3.

The G4 helicase DHX36 binds to Task3 mRNA and partially relieves the G4-mediated translation repression. (A) Endpoint RT-PCR of 3F-DHX36 mRNA targets isolated through FLAG immunoprecipitation in transfected HEK293 cells. (B) Western blotting of HEK-293 lysates co-transfected with each Task3 mutant 5′ UTR construct and either the G4 resolving helicase DHX36 or a non-G4 resolving mutant (DHX36 ΔDSM). (C) Quantification of endogenous and exogenous DHX36 expression for each co-transfection condition normalised to α-actin expression. (D) Quantification of Task3M1–3F expression normalised to α-actin expression. In each co-transfection condition, Task3M1-3F expression values for the Task3 5′ UTR mutant constructs were normalised to the ΔG4 control. (E) Western blotting of HEK293 lysates transfected with FL Task3–3F as well as either 3F-DHX36 WT or ΔDSM (Left) with exogenous FL Task3–3F expression quantification normalised to α-tubulin (Right). (F) Whole cell patch clamp recordings of HEK293 cells co-transfected with FL Task3–3F as well as either 3F-DHX36 WT or ΔDSM. Each data point represents an individual neuron. N = 4 biological replicates where error bars represent SEM. Statistical analyses were carried out by unpaired two-tailed t-tests, P-values shown. UT = untransfected.