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. Author manuscript; available in PMC: 2023 Dec 18.
Published in final edited form as: Cell Rep. 2023 Oct 25;42(11):113329. doi: 10.1016/j.celrep.2023.113329

Figure 6. PRMT5 mediates methylation of FoxO1 to alter its subcellular localization.

Figure 6.

(A) Western blot images showing subcellular fraction of control (MeOH) and Prmt5 KO (4-OH) myoblasts and cytoplasmic retention of FoxO1 in Prmt5 KO cells. KD, kilodalton size marker.

(B) Pax7 and FoxO1 immunofluorescence (left) of SCs on freshly isolated myofibers from WT and Prmt5MKO mice, along with quantification of FoxO1 subcellular location (right) (n = 5). Scale bar, 10 μm.

(C) Control (MeOH) and Prmt5 KO (4-OH) myoblasts were immunoprecipitated with SYM10 and PRMT5 antibody and blotted with FoxO1, PRMT5, and GAPDH antibodies. KD, kilodalton size marker.

(D and E) Quantification of methylated FoxO1 (D) and PRMT5-FoxO1 binding (E) (n = 3).

(F) Control (MeOH) and Prmt5 KO (4-OH) myoblasts were immunoprecipitated with SYM10 after subcellular fractionation and blotted with PRMT5, FoxO1, GAPDH, and histone 3 antibodies. KD, kilodalton size marker.

(G and H) Quantification of methylated FoxO1 in the nucleus (G) and cytoplasm (H) (n = 4).

(I and J) Proximity ligation assay (PLA) on control (MeOH) and Prmt5 KO (4-OH) myoblasts using SYM10 and FoxO1 (I), or PRMT5 and FoxO1 (J) antibody pairs.

The presence of red punta indicates potential protein modification or interaction. Scale bar, 10 μm. Values are expressed as mean ± SEM. *p < 0.05, **p < 0.01 by t test. See also Figure S6.