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. 2021 Sep 26;22(19):10371. doi: 10.3390/ijms221910371

Figure 3.

Figure 3

AK4 regulates the viability and proliferation of PASMCs both under normoxia and hypoxia. For all experiments, PASMCs were transfected with siRNA-targeting AK4 (siAK4) or non-targeting control (scrambled siRNA = scramble) followed by exposure to normoxia (NOX) or hypoxia (HOX, 1% O2) for the indicated time points. (A) Relative mRNA expression of AK4 in PASMCs after AK4 knockdown (siAK4), compared to corresponding control (scramble) under NOX or HOX for 24, 48, and 72 h; n = 3. (B) Representative Western blot for AK4 followed by densitometric quantification of relative expression in PASMCs after AK4 knockdown (siAK4), compared to corresponding control (scramble) under NOX or HOX for 24 h; n = 5. (C) PASMC apoptosis measured by pSIVA-IANBD probe after AK4 knockdown (siAK4), compared to corresponding control (scramble) under NOX or 24 h HOX; n = 6. (D) PASMC viability measured by alamarBlue assay after AK4 knockdown (siAK4), compared to corresponding control (scramble) under NOX or HOX for 76 h; n = 6. (E) PASMC proliferation assessed by BrdU incorporation assay after AK4 knockdown (siAK4), compared to corresponding control (scramble) under NOX or HOX for 72 h; n = 5. (F) Relative mRNA expression of Ki-67 in PASMCs after AK4 knockdown (siAK4) under normoxia (NOX) or hypoxia (HOX) for 72 h, compared to scramble; n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, two-way ANOVA followed by Bonferroni multiple comparisons test.