Figure 3. The effect of tungstate on JMJD1A and JARID1A protein levels in BEAS-2B and A549 cells, and the effect of tungstate on relative mRNA levels of JMJD1A and JARID1A in BEAS-2B.

BEAS-2B cells were exposed to 0, 1, 2.5 and 5 mM of Na₂WO₄ for 48-hours, and then cells were allowed to proliferate in the absence of tungstate for 48 hours, for a washout. Nuclear extracts were extracted after treatment and washout.

A, JMJD1A was detected using Western blotting. The same membrane was re-probed with Lamin A antibody as loading control.

The results were repeated in three independent experiments; one representative blot is shown here. Graphical representations of relative intensities are calculated by ImageJ.

B, JARID1A was detected using Western blotting. The same membrane was re-probed using Lamin A antibody as loading control.

The results were repeated in three independent experiments; one representative blot is shown here. Graphical representations of relative intensities are calculated by ImageJ.

C, A549 cells were exposed to 0, 2.5 and 5 mM of Na₂WO₄ for 48-hours, and then cells were allowed to proliferate in the absence of tungsten for 48 hours, for a washout. Nuclear extracts were extracted after treatment and washout. JARID1A was detected using Western blotting and the same membrane was re-probed using Lamin A antibody as loading control.

The results were repeated in three independent experiments; one representative blot is shown here. Graphical representations of relative intensities are calculated by ImageJ.

D, After 48-hour acute treatment of 2.5 and 5 mM Na₂WO₄, RT-qPCR of histone demethylases, JMJD1A and JARID1A, was evaluated.

Statistical significance was calculated using an unpaired, two-tailed t test

* p-value < 0.05

** p-value < 0.01

*** p-value < 0.001

Error bars represent Standard Deviation.