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. 2023 Oct 11;26(11):108168. doi: 10.1016/j.isci.2023.108168

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

MUC1-C represses mtDNA gene expression

(A) Expression of the indicated mtDNA genes in 3D vs. 2D BT-549 cells was determined by qRT-PCR. The results (mean ± SD of 3 determinations) are expressed as relative mRNA levels compared to that obtained for 2D cells (assigned a value of 1). Asterisks represent ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001.

(B and C) Expression of the indicated mtDNA genes in (B) BT-549/tet-MUC1shRNA 3D cells treated with DOX vs. vehicle for 10 days, and (C) BT-549 3D cells treated with 5 μM GO-203 vs. vehicle for 36 h was determined by qRT-PCR. The results (mean ± SD of 3 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). Asterisks represent ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001.

(D) Lysates from BT-549/tet-MUC1shRNA 3D cells treated with DOX vs. vehicle for 7 days (left) and BT-549 3D cells treated with 2.5 μM GO-203 vs. vehicle for 2 days (right) were immunoblotted with antibodies against the indicated proteins.

(E) BT-549/tet-MUC1shRNA 3D cells treated with DOX vs. vehicle for 4 days (left) and BT-549 3D cells treated with 5 μM GO-203 vs. vehicle for 2 days (right) were analyzed for ATP levels. The results (mean ± SD of 3 determinations) are expressed as relative ATP levels compared to that obtained for vehicle-treated cells (assigned a value of 1). Asterisks represent ∗∗p ≤ 0.01, ∗∗∗∗p ≤ 0.0001.

(F) BT-549/tet-MUC1shRNA 3D cells treated with DOX vs. vehicle for 5 days were analyzed for TFAM expression by qRT-PCR. The results (mean ± SD of 3 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). Asterisks represent ∗∗∗∗p ≤ 0.0001.

(G) Lysates from BT-549/tet-MUC1shRNA 3D cells treated with DOX vs. vehicle for 7 days (left) and BT-549 3D cells treated with 2.5 μM GO-203 vs. vehicle for 2 days (right) were immunoblotted with antibodies against the indicated proteins.

(H) Lysates from BT-549/tet-MYCshRNA 3D cells treated with vehicle or DOX were immunoblotted with antibodies against the indicated proteins.

(I) BT-549/tet-MUC1shRNA (left) and BT-549/tet-MYCshRNA (right) 3D cells treated with vehicle or DOX were analyzed for mTERF3 expression by qRT-PCR. The results (mean ± SD of 3 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). Asterisks represent ∗∗p ≤ 0.01, ∗∗∗∗p ≤ 0.0001.

(J) Lysates from BT-549/tet-MUC1shRNA and BT-549/tet-MYCshRNA 3D cells treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins.