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Investigating the Role of TLR2 and TLR4 in Palmitate-Induced INS-1 Cell Death

This study explores the effects of Toll-like receptors (TLR2 and TLR4) on palmitate-induced cell death in INS-1 cells. Using pipette-based electroporation, INS-1 cells were transfected with GFP to assess transfection efficiency. The protective effect of the calcineurin inhibitor deltametrin on cell viability was evaluated alongside phospho-JNK and IκBα levels through immunoblotting. DNA fragmentation was measured to quantify cell death. Results highlight the interaction between palmitate, TLRs, and deltametrin, with significant findings supporting potential therapeutic targets.

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Investigating the Role of TLR2 and TLR4 in Palmitate-Induced INS-1 Cell Death

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  1. Supplement 1 siRNA sequences of green fluorescent protein (GFP), toll-like receptor 2 (TLR2), and toll-like receptor 4 (TLR4)

  2. Supplement 2 B A a b Supplement 2. GFP-positive cells after pipette-based electroporation. After transfection of 1 g pEGFP-C1 DNA (Clontech) into 5x105 INS-1 cells using a pipette type-electroporator, the cells were further incubated for 48 h. The transfected cells were observed under phase contrast microscope (a) and fluorescent microscope (b) (A). Transfection efficiency was determined by counting GFP-positive fluorescent cells from total cells (B). Total Cells GFP-positive Cells

  3. Supplement 3 Primer sequences and reaction conditions for semi-quantitative RT-PCR

  4. Supplement 4 * * B DNA fragmentation (OD) A IB Del (M): 0 0.01 0.1 1 0 0.01 0.1 1 PA P-JNK 0 0.5 4 8 12 16 (h) T-JNK PA + Deltametrin actin Supplement 4. Protective effect of calcinurin inhibitor deltametrin on palmitate-induced INS-1 cell death and augmentation effect of deltametrin on the palmitate-induced TLR downstream signaling pathway. (A) INS-1 cells were treated with 0.4 mM palmitate and 0.1 M deltametrin for the indicated times. Levels of phospho-JNK and IB were determined by immunoblotting analysis of total lysates with anti-phospho-JNK and anti-IB antibodies. (B) INS-1 cells were treated with 0.4 mM palmitate for 16 h in the presence of different concentrations of deltametrin. Cell death was determined by measurement of DNA fragmentation using Cell Death Detection ELISA kit. Data represent mean + SE from three independent experiments. *P<0.05 vs. absorbance from untreated INS-1 cells.

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