Figure 1.

In Vitro Validation of shRNA Targeting DNM2

(A) Location of shRNA-targeting sequences in Dnm2 mRNA. Exons are depicted as boxes, and exons 10a, 10b, 12b, and 13b are alternative exons. Below, the protein domains encompass a GTPase domain, a middle and GTP effector (GED) domains forming the stalk, a pleckstrin homology (PH), and a proline-rich (PRD) domain. (B) Representative western blot from HEK293T cells co-transfected with two plasmids is shown: one encoding human DNM2 and the second expressing the different shRNA. DNM2 protein levels were determined by densitometry and standardized to GAPDH. (C) Endogenous Dnm2 mRNA levels in shRNA-treated C2C12 mouse myoblast cells were determined by qRT-PCR and standardized to Hprt. Cells were electroporated with shRNA ctrl or shDnm2 (B, C, F, I, or J). n = 3 biological replicates per each group. Data represent an average of three independent experiments ± SEM. *p < 0.05; **p < 0.01 for shDnm2-treated versus shRNA ctrl-treated cells (ANOVA test).