siRNA treatment

MDA-MB-231 cells were transfected with a negative siRNA control (4390843; Life Technologies) and with NUCB1 or MMP2 siRNA at a final concentration of 5 nM using RNAiMax (Life Technologies) according to the manufacturer’s instructions. siRNA targeting firefly luciferase mRNA (D-001210-02-20; Dharmacon) was used as a negative control in primary human macrophages, which were transfected with NUCB1 or MMP2 siRNA at a final concentration of 100 nM. Knockdowns were achieved 72 h after transfection using the Viromer Blue Kit according to the manufacturer’s instructions. Reduction of targeted proteins was validated by WB of respective cell lysates.