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Puromycin incorporation assay (SUnSET assay)
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SUnSET assay was performed as per manufacturer’s recommendations (Kerafast). Cells were incubated with puromycin (normal cells (non-infected): 1 µg/mL, infected cells: 3 µg/mL) for 30 min. Post incubation, cells were washed with ice cold PBS and lysed using RIPA lysis buffer. Equal quantity of protein lysates was separated on SDS-PAGE and probed with anti-puromycin antibody. Signals were normalized with probing GAPDH (loading control).
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