Fig. 6

The reduction of protein translation promotes Vegfa expression independent of Hif1α. a Western blot analysis of the p-Eif2α, Eif2α, and α-tubulin of B16-Red transplant microtumor at avascular (about 12 h)and first vessel stage (about 24 h) (n = 50). b Western blot analysis of the p-Eif2α, Eif2α, and α-tubulin of B16-Red tumor spheroids at 0, 12 and 24 h. c RT-qPCR quantification of Vegfa expression in B16-Red tumor spheroids at 0, 12 and 24 h. d Western blot analysis of the p-Eif2α, Eif2α, and α-tubulin of puromycin (2 μg/mL) treated B16-Red cells at 0, 1 and 3 h. e–h RT-qPCR quantification of Vegfa expression in puromycin (4 μg/mL) treated B16-Red cells or in puromycin (1 μg/mL) treated H460, SKOV3 and A2780s cells for 24 h. i-l RT-qPCR quantification of Vegfa expression in cycloheximide (800 μM) treated B16-Red and H460 or in cycloheximide (200 μM) treated SKOV3 and A2780s cells for 24 h. m RT-qPCR quantification of Vegfa expression in B16-Red cells. Untreated (Control), negative control or Hif1α siRNA transfected for 48 h (siNC or siHif1α), puromycin (2 μg/mL) treated for 24 h (Puromycin), and puromycin (2 μg/mL) treated for 24 h and Hif1α siRNA transfected for 48 h (siHif1α + Puromycin). Control, Con. Puromycin, Puro. Cycloheximide, CHX. Unpaired t-test, *P < 0.05, ** P < 0.01, *** P < 0.001 (n = 3)