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Protein extraction and western blotting For protein isolation, hDFs, iPSCs and MNPs were washed twice with ice-cold 1X phosphate saline buffer (1X PBS) and lysed with an extraction buffer containing 50 mM HEPES (Thermo Fisher Scientific), 150 mM NaCl (Applichem), 1 mM EDTA (pH8) (Applichem), 1 mM EGTA(pH8), 10% glycerol (Romil Pure Chemistry), and 1% Triton-X-100 (Applichem) complemented with protease (Sigma-Aldrich) and phosphatase inhibitors (PhosSTOP, Roche), according to the manufacturers protocol

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Treatment of slow-transit constipation with sacral nerve modulation

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Each group contributes to the area's rich cultural tapestry through unique traditions, languages, and customs