Fig 1: CHL1 mediates DDP and PTX sensitivity by inhibiting Akt activity. (A) Western blotting was performed to detect the expression of p-Akt and total Akt in CHL-silenced and -restored cell models. *P<0.05 vs. scramble or vector. (B) MTT assays were performed to detect cell survival rates of A549 cells treated with CHL1 siRNA and Akt inhibitor SC66. *P<0.05. (C) Colony formation assays were performed in A549 cells treated with CHL1 siRNA and the Akt inhibitor SC66 in the presence of DDP (1.5 µg/ml) or PTX (35 ng/ml). *P<0.05 vs. si-CHL1. (D) Apoptosis were measured in A549 cells treated with CHL1 siRNA and Akt inhibitor SC66 in the presence of DDP (1.5 µg/ml) and PTX (35 ng/ml). *P<0.05 vs. si-CHL1. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel; si, small interfering; p-, phosphorylated.
Fig 2: CHL1-knockdown increases A549 cell resistance to DDP and PTX. (A) Western blotting was performed to validate the efficiency of transfection with CHL1 siRNAs. *P<0.05 vs. scramble. MTT assays were performed to determine the survival rate of CHL1-knockdown A549 cells treated with (B) 0–2.5 µg/ml DDP or (C) 0–50 ng/ml DDP. (D) Colony formation assay of A549 cells transfected with CHL1 siRNA in the presence or absence of 1.5 µg/ml DDP or 35 ng/ml PTX. (E) Flow cytometry analysis was used to detect apoptosis in A549 cells transfected with CHL1 siRNA in the presence or absence of 1.5 µg/ml DDP or 35 ng/ml PTX. *P<0.05, ***P<0.001. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel; si, small interfering.
Fig 3: Overexpression of CHL1 increases the sensitivity of resistant A549 cells to DDP and PTX. (A) Western blotting was performed to detect CHL1 expression in A549/DDP and A549/PTX cells transfected with CHL1 expression plasmids. *P<0.05 vs. vector. Effect of CHL1 overexpression on resistant A549 cell survival rate when treated with (B) 0–10 µg/ml DDP or (C) 0–250 ng/ml PTX, as determined by MTT assay. (D) Colony formation assays demonstrated the number of colonies of resistant A549 cells transfected with CHL1 expression plasmids in the presence or absence of 8 µg/ml DDP or 160 ng/ml PTX. (E) Flow cytometry analysis was performed to assess apoptosis in resistant A549 cells transfected with CHL1 expression plasmids in the presence or absence of 8 µg/ml DDP or 160 ng/ml PTX. CHL1 overexpression enhanced chemosensitivity of A549/DDP cells to DDP in vivo, which was demonstrated by the effect of DDP treatment or CHL1 overexpression on the (F) growth and (G) weight of xenografts derived from A549/DDP cells. *P<0.05, **P<0.01. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel.
Fig 4: CHL1 is downregulated in DDP and PTX-resistant A549 cells. (A) Cell survival of A549 and A549-resistant cells (A549/DDP and A549/PTX) treated with increasing concentrations of DDP and PTX, as assessed by MTT assay. (B) The IC50 values of DDP in A549/DDP and A549 cells, and the IC50 values of PTX in A549/PTX and A549 cells. *P<0.05 vs. A549 cells. (C) Western blotting demonstrated the expression of drug resistance-related proteins MDR1, MRP and LRP in A549 cells and A549-resistant cells (A549/DDP and A549/PTX). *P<0.05 vs. A549 cells. The protein and mRNA expression levels of CHL1 in A549 cells and A549-resistant cells (A549/DDP and A549/PTX) were analysed by (D) western blotting and (E) reverse transcription-quantitative PCR, respectively. *P<0.05 vs. A549 cells. (F) The mRNA expression of CHL1 in H460 and H460/DDP cells in the GSE21656 dataset. *P<0.05 vs. H460 cells. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel; MDR1, multi-drug resistance gene 1; MRP, multidrug resistance-associated protein; LRP, low-density lipoprotein receptor-related protein; IC50, half maximal inhibitory concentration.
Supplier Page from Proteintech Group Inc for CHL1 antibody