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. 2023 Oct 5;26(11):108142. doi: 10.1016/j.isci.2023.108142

Figure 4.

Figure 4

Expression of ALDH1A1 with Khib modification regulated by HDAC2/3

(A) Flag-tagged ALDH1A1 was co-transfected with vector or HA-tagged HDAC2/HDAC3 respectively as indicated.

(B and C) Flag-tagged ALDH1A1 was overexpressed in T24 cells, and the Khib of ALDH1A1 was measured by knocking down HDAC2 and HDAC3.

(D and E) Endogenous ALDH1A1 was IPed from T24 cells. Rabbit IgG was used as a control.

(F and G) Western blotting analysis of HDAC2/3 and ALDH1A1 in HDAC2/3 overexpressed and knocking down T24 cells.

(H) T24 cells were either untreated or treated with TSA for different lengths of time as mentioned earlier, meanwhile, the cells were cultured with cycloheximide (CHX) and harvested at indicated time points. Then, immunoblotting against ALDH1A1 and α-actinin control was performed.

(I) MG132 was added into the culture medium without or with TSA in T24 cells, as indicated to determine the ubiquitination of ALDH1A1.For cell experiments, each experiment was performed at least three times.