Fig 1: miR-210 attenuates the hypoxia-induced increase in DNA fragmentation through the inhibition of GSK3ß kinase activity. (A) Quantitative TUNEL assay determining apoptotic DNA fragmentation, expressed as experimental blank-corrected absorbances (O.D) measured at ?450 (450 nm). (B) Qualitative Western blot depicting the processing of the DFF45 into cleaved-DFF45 (p11 fragment) as a surrogate measure of caspase-3-induced DFF40 endonuclease activity. (C) Quantitative ELISA determining the levels of cleaved-DFF45 (p11 fragment), expressed as experimental blank-corrected absorbance (O.D) measured at ?570 (570 nm). (D) Quantitative DFF40 endonuclease activity assay, expressed as experimental blank-corrected absorbances (O.D) measured at ?405 (405 nm). miR-210 expression levels in the respective cell lysates were determined by the miR-210 hybridization immunoassay (as described in Section 4.3) and are reported in Supplementary Figure S2A. The validation of the ectopic expression of the HA-tagged ca-GSK3ß-S9A mutant in the pertinent experimental groups was performed by ELISA immunoassay and is reported in Supplementary Figure S2B). GSK3ß kinase activity (as described in Section 4.4) was measured in all experimental groups to corroborate and validate the translative effects of the ectopic expression of the ca-GSK3ß-S9A mutant (reported in Supplementary Figure S2C). All data are expressed as mean ± S.D from three technical replicates for each of the four biological replicates belonging to each experimental group (n = 4). ** p = 0.01; **** p = 0.0001; ns: not significant (p > 0.05). OE: miR-210 overexpression; O.D: optical density; S.D: standard deviation.