Fig. 6: PCK1 may play a significant role in Akt activation in uveal melanoma cells.

The 92.1 cells were engineered to stably express the lentiviral PCK1 shRNA (“shPCK1-seq-1” or “shPCK1-seq-2”, the lentiviral control shRNA (“shC”) (A), the lentiviral CRISPR/Cas9-PCK1-KO construct (“koPCK1”), the CRISPR/Cas9-control construct (“Cas9-C”) (B), the lentivirus-packed PCK1-expressing construct (“oePCK1”) or the empty vector (“Vec”) (C), p-Akt (at Ser-473) and Akt1 expression was tested. The shPCK1-seq-2-expressing 92.1 cells were subjected to stable transduction with a constitutively-active mutant Akt1 bearing the S473D mutation (“caAkt1”) or an empty vector (“Vec”). The expression levels of the specified proteins were shown (D). An equal number of the aforementioned 92.1 cells were cultured for specific durations, cell proliferation, migration, and apoptosis were assessed using EdU-nuclei staining (E), “Transwell” assays (F), and TUNEL-nuclei staining (G), respectively. The numerical values are presented as the mean ± standard deviation (SD). “Pare” signifies the parental control cells. *Indicates statistical significance (P < 0.05) when compared to “shC”/”Cas9-C”/”Vec” cells (A–C). ^#P < 0.05 (D-G). “N. S.” denotes a lack of statistical difference (P > 0.05). The experiments depicted in this figure were replicated five times (n = 5, biological repeats), consistently yielding similar results. The scale bar corresponds to 100 μm.