GFP/RFP intensity quantification

GFP/RFP intensities before and after the addition of inhibitor (Binucleine two or BI 2536) were quantified to estimate the effects of inhibitor on CS-TIPs patterning and signaling. Identical regions were drawn over the GFP/RFP puncta to measure the fluorescence intensity. The local background was estimated by placing the same region to a nearby place. Background intensity values were subtracted from GFP/RFP values to estimate the actual GFP/RFP fluorescence intensity. Histograms and dot plots were generated using Prism software (Graphpad, CA, USA), and figures were assembled using Adobe Illustrator software (Adobe, CA, USA). Statistical analysis was performed using Prism software (Graphpad, CA, USA).