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Transwell chambers were assayed for cell invasion ability by applying 100 μL of DMEM-diluted Matrigel gel (3:7 dilution) to the chambers to be checked and incubated at 37 °C for 30 min to form a gel, then pre-treated MIA PaCa-2 and PANC-1 cells were resuspended with DMEM and spread in small chambers at 50000/well, and the lower chamber was placed in 20% FBS DMEM, incubated at 37°C 5% CO 2 for 48 hours

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Common indicators include: Fatigue and Weakness: Persistent tiredness and low energy are often the first signs of low vitamin D