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. 2021 Feb 4;16(2):e0246531. doi: 10.1371/journal.pone.0246531

Fig 2. HIF-1alpha protein levels in PDI-overexpressing cells.

Fig 2

(A-F) PDI was overexpressed in Hep3B cells (A and D), HEK293T cells (B and E), and HeLa cells (C and F), and cells were cultured for 6 h under hypoxic conditions. Proteins (3 or 15 μg) were separated by SDS-PAGE and immunoblotting was performed with an anti-PDI, -HIF-1alpha, and -beta-actin antibody (1:1000 dilution). (G) Hep3B cells, HEK293T cells, and HeLa cells were cultured for 6 h under hypoxic conditions. The expression of PDI was evaluated by immunoblotting with an anti-PDI antibody. (H) The PDI/3×FLAG-pcDNA4 vector (6 or 12 μg) was transfected into Hep3B cells, and cells were cultured for 6 h under hypoxic conditions. Proteins (3 μg) were separated by SDS-PAGE, and immunoblotting was performed with an anti-FLAG tag antibody (1:1000 dilution). (I) PDI/pcDNA 3.1 (+) and si-PDI were transfected into Hep3B cells, cells were cultured for 6 h under hypoxic conditions, and immunoblotting was performed. The PDI or HIF-1alpha protein levels of mock cells (A-F and H), or si-ctrl cells (I) under hypoxic conditions were set at 1.0. Values are expressed as the mean ± S.D. (error bars) of three different plates.   N, Normoxia. * p<0.05; ** p<0.01, significantly different from mock or ctrl cells (A-G) or si-ctrl cells (I) under hypoxic conditions.