Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Effect of BITC on CNOT2 and pSTAT3 and their binding in SK-Hep1 and Huh7 cells. ( A ) Overexpression of CNOT2 in liver cancer patients implies poor survival rate along with original regression ratio with STAT3. ( B ) Effect of BITC on CNOT2 and pSTAT3 in SK-Hep1 and Huh7 cells. Band intensities were quantified and normalized to β-actin. ( C ) Effect of BITC on the binding between CNOT2 and STAT3 or c-Myc in SK-Hep1 cells. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: Binding Assay, Over Expression

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Depletion of STAT3 or CNOT2 enhances apoptotic effect of BITC in SK-Hep1 cells. ( A ) Effect of STAT3 or CNOT2 depletion in BITC treated SK-Hep1 cells. ( B ) Effect of STAT3 depletion on pro-PARP and pro-caspase3 in BITC treated SK-Hep1 cells. ( C ) Effect of CNOT2 depletion on pro-PARP and pro-caspase3 in BITC treated SK-Hep1 cells. Band intensities were quantified and normalized to β-actin. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques:

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Effect of BITC on Warburg effect in SK-Hep1 and Huh7 cells. ( A ) Effect of BITC on Warburg effect proteins in SK-Hep1 and Huh7 cells. ( B ) Effect of BITC on LDH production in SK-Hep1 and Huh7 cells. STAT3 or CNOT2 depletion vs untreated control. ***p < 0.001 vs untreated control. ( C ) Effect of BITC on glucose consumption in SK-Hep1 and Huh7 cells. *p < 0.05, ***p < 0.001 vs untreated control. ( D ) Effect of pyruvate treatment or overexpression of CNOT2 or c-Myc on apoptosis and glycolysis proteins in BITC treated SK-Hep1 cells. (E) Effect of siSTAT3 or/and siCNOT2 on HK2, PKM2 and LDH in SK-Hep1 cells transfected with or without c-Myc overexpression plasmid. Cells were co-transfected with control vector, siSTAT3, siCNOT2 and siCNOT2 + c-Myc OE (overexpression) plasmids as indicated in each lane. Band intensities were quantified and normalized to β-actin. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: Control, Over Expression, Transfection, Plasmid Preparation

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Scheme on mechanism of BITC via CNOT2/c-Myc/STAT3 signaling in HCCs.

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: