Journal: Oncology Reports

Article Title: SOX2 regulates paclitaxel resistance of A549 non-small cell lung cancer cells via promoting transcription of ClC-3

doi: 10.3892/or.2022.8396

Figure Lengend Snippet: Primer sequences.

Article Snippet: The primary antibodies used were: ClC-3 (1:1,000; cat. no. 13359; Cell Signaling Technology, Inc.), SOX2 (1:1,000; cat. no. 23064; Cell Signaling Technology, Inc.), octamer-binding transcription factor 4 (OCT4; 1:1,000; cat. no. 2750; Cell Signaling Technology, Inc.), NANOG (1:1,000; cat. no. 4903; Cell Signaling Technology, Inc.), KLF transcription factor 4 (KLF4; 1:1,000; cat. no. 4038; Cell Signaling Technology, Inc.), multidrug resistance mutation 1 (MDR1; 1:1,000; cat. no. 13342; Cell Signaling Technology, Inc.), ATP binding cassette subfamily C member 2 (ABCC2; 1:1,000; cat. no. 4446; Cell Signaling Technology, Inc.), ATP binding cassette subfamily C member 10 (ABCC10; 1:1,000; cat. no. ab69296; Abcam), GAPDH (1:10,000; cat. no. ARG65680; Arigo Biolaboratories Corp.) and tubulin (1:10,000; cat. no. ARG65693; Arigo Biolaboratories Corp.).

Techniques:

Journal: Oncology Reports

Article Title: SOX2 regulates paclitaxel resistance of A549 non-small cell lung cancer cells via promoting transcription of ClC-3

doi: 10.3892/or.2022.8396

Figure Lengend Snippet: Higher levels of SOX2 confer PTX resistance in A549 non-small cell lung cancer cells. (A) mRNA expression levels of stemness-related genes were measured by reverse transcription-quantitative PCR in A549 and A549-PTX cells. (B) (Left) Protein expression levels of NANOG, KLF4, OCT4 and SOX2 were examined by western blotting in A549 and A549-PTX cells. (Right) Protein expression was semi-quantified using ImageJ. (C) Protein expression levels of SOX2 were examined by western blotting in A549-PTX cells transfected with siNC or SOX2 siRNA (siSOX2-1, siSOX2-2 and siSOX2-3). (D) Viability was examined using CCK-8 assays in A549-PTX cells transfected with siSOX2 or its control and treated with PTX for 48 h. (E) (Left) Protein expression levels of MDR1, ABCC10, ABCC2 and SOX2 were examined by western blotting in A549-PTX cells transfected with siSOX2 or its control. (Right) Protein expression was semi-quantified using ImageJ. (F) Protein expression levels of SOX2 were examined by western blotting in A549 cells overexpressing SOX2 or its vector control. (G) Viability was examined using CCK-8 assays in A549, A549-PTX and SOX2-overexpressing A549 cells treated with PTX for 48 h. (H) (Left) Protein expression levels of MDR1, ABCC10 and ABCC2 were examined by western blotting in A549 cells overexpressing SOX2 or its vector control. (Right) Protein expression was semi-quantified using ImageJ. GAPDH or Tubulin were used as a loading control in western blotting. All data are presented as the mean ± standard deviation. *P<0.05, ***P<0.001. ns, not significant. Relative, vs. respective control. SOX2, SRY-box transcription factor 2; PTX, paclitaxel; A549-PTX cells, PTX-resistant A549 non-small cell lung cancer cells; KLF4, Krüppel like factor 4; OCT4, octamer-binding transcription factor 4; siRNA/si, small interfering RNA; siNC, control siRNA; CCK-8, Cell Counting Kit-8; MDR1, multidrug resistance mutation 1; ABCC10, ATP binding cassette subfamily C member 10; ABCC2, ATP binding cassette subfamily C member 2; Ctl, control.

Article Snippet: The primary antibodies used were: ClC-3 (1:1,000; cat. no. 13359; Cell Signaling Technology, Inc.), SOX2 (1:1,000; cat. no. 23064; Cell Signaling Technology, Inc.), octamer-binding transcription factor 4 (OCT4; 1:1,000; cat. no. 2750; Cell Signaling Technology, Inc.), NANOG (1:1,000; cat. no. 4903; Cell Signaling Technology, Inc.), KLF transcription factor 4 (KLF4; 1:1,000; cat. no. 4038; Cell Signaling Technology, Inc.), multidrug resistance mutation 1 (MDR1; 1:1,000; cat. no. 13342; Cell Signaling Technology, Inc.), ATP binding cassette subfamily C member 2 (ABCC2; 1:1,000; cat. no. 4446; Cell Signaling Technology, Inc.), ATP binding cassette subfamily C member 10 (ABCC10; 1:1,000; cat. no. ab69296; Abcam), GAPDH (1:10,000; cat. no. ARG65680; Arigo Biolaboratories Corp.) and tubulin (1:10,000; cat. no. ARG65693; Arigo Biolaboratories Corp.).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Transfection, CCK-8 Assay, Control, Plasmid Preparation, Standard Deviation, Binding Assay, Small Interfering RNA, Cell Counting, Mutagenesis