A) Confocal reconstruction of MDA-MB-468 cells invading into HANDBF/MMPx hydrogels with medium gradients of EGF555 and Alexa546 B) Normalized invasion distance of MDA-MB-468 cells in medium gradients of Alexa546 and EGF555 and non-patterned regions of the HANDBF/MMPx hydrogel (day 6, n=3, mean + standard deviation). EGF555 significantly decreased MDA-MB-468 invasion distance compared to Alexa546 on both the gradient and non-patterned regions (****p<0.0001, two-way ANOVA). C) Normalized cell number of MDA-MB-468 cells in medium gradients of Alexa546 and EGF555 and non-patterned regions of the HANDBF/MMPx hydrogel (day 6, n=3, mean + standard deviation). EGF555 significantly decreased MDA-MB-468 cell number compared to Alexa546 on both the gradient and non-patterned regions (****p<0.0001, two-way ANOVA). D) Average cell diameter of MDA-MB-468 cells in medium gradients of Alexa546 and EGF555 and non-patterned regions of the HANDBF/MMPx hydrogel (day 6, n=3, mean + standard deviation). EGF555 significantly increased MDA-MB-468 cell diameter compared to Alexa546 on both the gradient and non-patterned regions (***p<0.001, two-way ANOVA). E) Representative images showing MDA-MB-468 cells with larger diameters when cultured on EGF555 gradients compared to the control Alexa546 gradients.