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. 2023 Apr 4;120(15):e2207898120. doi: 10.1073/pnas.2207898120

Fig. 1.

Fig. 1.

Overexpression of miR-183 down-regulates eIF2Bδ protein. (A) Scheme for identification of miRs altered in expression in cancer stem cell populations from immortalized and progressively transformed breast cancer (BC) cell populations. MMs, mammospheres. (B) Heat map comparing the relative expression of miRs from the breast CSC population and the parental cell line. (C) Alignment of miR-183 seed sequence to the coding region of eIF2Bδ mRNA, present in each of the eIF2Bδ variants. (D) Representative immunoblot from cell lines expressing miR-Scr nonsilencing control or miR-183, by transient transfection or stable expression. eEF2, invariant loading control. n = 3. (E) Representative immunoblot of ezrin protein levels in cells expressing Dox-induced miR-Scr, miR-183, or shRNA to eIF2Bδ (sh-2Bδ). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), invariant loading control. n = 3. (F) Representative immunoblot of eIF2Bδ and GAPDH control protein levels in cells expressing a scrambled sequence (Zip-Scr) or antagomir to miR-183 (Zip-183) in cells stably expressing miR-Scr or miR-183. n = 3. (G) Protein synthesis rate in cells expressing miR-Scr or miR-183. *P < 0.05, ± SEM by two-tailed unpaired t test. n = 3.