Fig. 4
From: Sec23a inhibits the self-renewal of melanoma cancer stem cells via inactivation of ER-phagy
Sec23a inhibits the stemness of M14-SE cells via inhibition of autophagy. a Western blot analysis for LC3I/II and P62 in M14-SE cells with Sec23a interference or overexpression. b Representatives of LC3B-positive puncta images were shown in M14-SE cells with Sec23a interference or overexpression, bar = 20 μm. c Transmission electron microscopy of M14-SE cells with Sec23a interference or overexpression. Autolysosomes indicated by arrowheads, bar = 5 μm. d Mean number of detectable autolysosomes in each tumor cell, counted on transmission electron microscopy images. e and f RT-qPCR and Western blot analysis to confirm effective Atg5 interference in M14-SE-shSec23a cells. g Analysis of mRNA expression of Aldh1, CD133, Nanog, Oct4 and Sox2 in M14-SE-shSec23a cells with Atg5 interference. h–j. Clonogenic spheroid formation rate, morphology and average spheroid diameter of M14-SE-shSec23a cells with Atg5 interference in the single-cell cloning assay. k Sserial spheroid formation assay of M14-SE-shSec23a cells with Atg5 interference, bar = 200 µm. l Tumors resected from nude mice inoculated with M14-SE-shSec23a cells with Atg5 interference, bar = 1 cm. (m and n). Tumor volume and weight, **P < 0.01, ***P < 0.001