Fig 1: Stathmin-2 localizes to secretory granules in aTC1-6 cells. aTC1-6 cells were immunostained using primary antibodies against glucagon (GCG, green) and stathmin-2 (Stmn2, red). DAPI (blue) indicates the nucleus in the merged image. Resolution of the images was extended by applying Nyquist XY scan and then 2D- Deconvolution in NIS Elements image analysis software. Images are representative of four biological replicates with 3 technical replicates each. (A) Areas of yellow in the merged image show colocalization of glucagon and Stmn2. (B) Linear regression analysis of binary intensities of glucagon and Stmn2 predicts a significant (p < 0.001) correlation. Each value represents mean intensities of 5-7 cells. The secretion of both glucagon and Stmn2 (C) was significantly increased after KCl stimulation (KCl Stim)for 15 min. (D) Cell Stmn2 and glucagon levels show reduction following KCl stimulation (KCl Stim). Values are expressed as mean ± SEM (n = 5). *p < 0.05. Stmn2 colocalizes with the secretory granule proteins ChgA (E) and VAMP2 (F), as indicated by yellow punctate staining. (G) The extent of colocalization was analyzed by Pearson correlation coefficient for Stmn2 with ChgA or with VAMP2.