Skip to main content
. 2024 Oct 22;12:RP88318. doi: 10.7554/eLife.88318

Figure 3. Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) promotes the activation of mTORC1 pathway in human and mouse cells.

Figure 3.

(A) HEK293 cells treated with DYRK1A short hairpin RNA (shRNA) or control shRNA were serum starved for 12 hr before being activated with serum for the indicated times. Cells were then harvested, lysates, and probed with the indicated antibodies. Actin was used as the loading control. (B, C) Quantification of proteins in (A), levels of pS6K (T389), S6K, pS6 (pS235/236), and S6 were quantified using Image J software and the ratio of pS6K/S6K and pS6/S6 were plotted (n=3 biological replicates). (D) NIH3T3 cells were treated with sgRNA-targeting Dyrk1a or non-targeting control and selected for four days with Puromycin before harvesting. Lysates were probed with indicated antibodies. (E, F) Quantification of proteins in (D), levels of pS6K (T389), S6K, pS6 (pS235/236), and S6 were quantified (as described for B and C) and ratios were plotted (n=3 biological replicates). Student’s t-tests were done to compare samples. p-value = *p<0.05.

Figure 3—source data 1. Uncropped blots for Figure 3A.
Figure 3—source data 2. Raw data showing phosphorylation status of S6k and S6 after knockdown of DYRK1A for Figure 3A.
Figure 3—source data 3. Uncropped blots for Figure 3D.
Figure 3—source data 4. Raw data showing phosphorylation status of S6K and S6 after CRISPR knockout of Dyrk1a in NIH3T3 cells for Figure 3D.