Fig. 3.

Alterated linc01296 impacted the chemoresistance of CRC cells. a 5-FU resistant CRC cells were treated with different concentration of 5-FU, and the absorbance was measured at 450 nm. b The IC₅₀ values were calculated in control cells, siSCR group and silinc01296 group. Downregulation of linc01296 decreased IC₅₀ values in 5-FU resistant CRC cells. c Colony formation assay was used to detect the colony formation of CRC cells in response to 5-FU. d Flow cytometry was used to determine the apoptosis rate of different treated CRC cells. Knockdown linc01296 promoted CRC cells survival in response to 5-FU. e JC-1 assay was carried out to determine the mitochondrial membrane potential variation. Green fluorescence: the monomer, red fluorescence: the J-aggregates, orange fluorescence: merged photo. f TUNEL assay also confirmed the incidence of apoptosis by knocking down linc01296. g The molecular expression of key caspase-dependent apoptosis was determined by western blot. h CRC cells with high linc01296 level showed enhanced chemoresistance to 5-FU. i IC₅₀ values were increased in the cells transfected with linc01296. j Linc01296 facilitated more colony numbers than the vector in response to 5-FU. k More resistance to 5-FU was shown in CRC cells with high linc01296 level. Low apoptosis rate was calculated inlinc01296 transfected cells. l JC-1 immunofluorescence staining showed high mitochondrial membrane potential in HCT-8 transfected with linc01296. Green fluorescence: the monomer, red fluorescence: the J-aggregates, orange fluorescence: merged photo. m TUNEL depicted lower apoptosis occurrence in linc01296 transfected HCT-8 cells than the vector in response to 5-FU. (N) Caspase3, cleaved caspase3, PARP and cleaved PARP levels were analyzed by western blot. Data are the means ± SD of triplicate determinants (*P < 0.05)