Alizarin previous studies have shown that intramolecular interaction between ABD and MTBD of Shot prevents its interaction with actin. A similar mechanism may thus also affect the topogenic fate of BPAG1a/b. In line with our findings, Young et al. observed a strong perinuclear staining in C2C12 myoblasts using an antiserum directed against a peptide sequence specific to the N-terminal portion of the BPAG1a2/b2 isoforms. These data imply that BPAG1a2/b2 specifically contribute to the perinuclear staining that we observed in C2.7 myoblasts using R18024 antiserum. In contrast to our data, Young et al. also described co-localization of BPAG1a2/b2 with actin stress fibers in the cell center of 5-10% of C2C12 myoblasts as well as a nuclear staining of BPAG1a2/b2 in all cells. Since the R18024 antiserum used in our study binds to the SR region and should hence recognize all N-terminal isoforms of BPAG1a/b, the reasons for these staining differences remain unclear. Finally, in contrast to MACF1a, which co-localizes with focal adhesions in keratinocytes, BPAG1a/b did not co-localize with vinculin in FAs of C2.7 cells. Finally, as assessed by immunofluorescence, there was no difference in the depolymerisation of the MT network between control and BPAG1 knockdown cells upon treatment with nocodazole nor in the repolymerization of MTs after washing out nocodazole. Our findings are in line with previous studies showing that knockdown of BPAG1 in HFFF2 fibroblasts does not lead to any major change in the dynamics or organization of MTs. Since myoblasts and fibroblasts are structurally similar, and fibroblast Peramivir Trihydrate conversion to myogenic cells has been reported, it is very unlikely that BPAG1a/b serve as MT stabilizing proteins in these cell types. Finally, by immunofluorescence analysis, we examined the staining pattern of the actin network in BPAG1-knockdown myoblasts using phalloidin. When compared to control cells, BPAG1 knockdown had no impact on the MF network in transfected cells. Therefore, BPAG1a/b do not seem essential for either the stability of MTs or the organization of MT and MF networks in C2.7 myoblasts.