Research Paper Volume 14, Issue 22 pp 9149—9166

Figure 4. LINC00629 interacted with Mcl1. (A, B) UM-SCC6 cells with or without LINC00629 knockdown were pretreated with 40 μM for 33 h and were then treated with 10 μM CHX for the indicated times. The expression level of Mcl1 was determined by Western blotting. The results represent three independent experiments; ***p<0.001. (C, D) HA-Ub was transfected into Cal-27 and UM-SCC6 cells with or without LINC00629 knockdown. The cells were treated with MG132 for 8 h before collection. The whole-cell lysate was subjected to immunoprecipitation with an anti-Mcl1 antibody and Western blotting with an anti-HA antibody to detect ubiquitylated Mcl1. (E) The anti-Mcl1 antibody was used to coprecipitate LINC00629 in whole-cell lysates of UM-SCC6 cells treated with or without apigenin. (F) Biotin-labeled LINC00629 or antisense RNA was pulled down with Mcl1 in whole-cell lysates of UM-SCC6 cells. (G) Schematic illustration of the division of LINC00629 into three fragments corresponding to individual exons of the LINC00629 gene (E1, E2 and E3) along with the corresponding truncated bodies used. (H) Biotin-labeled LINC00629, truncated bodies or antisense RNA were pulled down with Mcl1 in whole-cell lysates of UM-SCC6 cells.