IL-1β expression levels in cell culture were determined using an ELISA kit (Enter life, UK). Cells were seeded in six-well plates at a density of 5 × 105 cells/well. Cell stimulation (normal, hyperosmolarity, CMC, α-MSH, CMC + α-MSH) was performed as above. After 24 h, cell supernatants were collected for ELISA analysis. ELISA procedures were performed in accordance with the manufacturer’s instructions.

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