Journal: Cell death & disease

Article Title: Transcription factor Sp1 transcriptionally enhances GSDME expression for pyroptosis.

doi: 10.1038/s41419-024-06455-6

Figure Lengend Snippet: Fig. 5 Sp1 is dispensable for other necrosis-associated gene expressions. A Western blotting analysis of lysates from Huh7 cells treated with indicated amount of plicamycin shows the protein expression levels of caspase 3, caspase 8, NINJ1, and β-actin. B, C qPCR analysis of caspase 3 and caspase 8 mRNA levels from Huh7 cells treated with the indicated amount of plicamycin, normalized by β-actin. D Western blotting analysis of lysates from iBMDM cells transfected with scramble/Sp1 shRNA. The protein expression levels of GSDMD and β-actin were detected. E, F iBMDM cells were treated with peretinoin for 24 h, followed by LPS and nigericin (Nig) stimulation for the indicated times. Cell viability and LDH release were determined, respectively. G Western blotting analysis of lysates from HT-29 cells transfected by scramble/ Sp1 shRNA shows the protein expression levels of human RIP3, MLKL, and β-actin. H, I HT-29 and MEF cells were treated with peretinoin for 24 h, followed by TNF, Smac-mimetic, and Z-VAD-FMK stimulation at the indicated times. Cell viability was determined.

Article Snippet: HT-29 and MEF cells were treated with 10 ng/ mL TNF (Abcam), 100 nM Smac-mimetic (MedChemExpress), and 5 μM ZVAD-FMK (Abcam) to induce necroptosis, and iBMDM cells were treated by 1 μg/mL LPS (Sigma), and 20 μM nigericin (MedChemExpress) to induce GSDMD-mediated pyroptosis.

Techniques: Western Blot, Expressing, Transfection, shRNA