NK Cell Perforin Expression Analysis

NK cells were treated with 50 nM CMA or DMSO for three hours at 37°C. Cells were lysed as previously described (Fasbender et al., 2017 (link)) and analyzed by Western blotting using the following antibodies: anti-human perforin (Pf-344; MabTech), β-actin (4967; Cell Signaling), and anti-CD107a (H4A3; BD Biosciences).

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Prager I., Liesche C., van Ooijen H., Urlaub D., Verron Q., Sandström N., Fasbender F., Claus M., Eils R., Beaudouin J., Önfelt B, & Watzl C. (2019). NK cells switch from granzyme B to death receptor–mediated cytotoxicity during serial killing. The Journal of Experimental Medicine, 216(9), 2113-2127.

Publication 2019

Pf-344; β-actin anti-CD107a (H4A3;

Actin Antibodies Cells Dmso Human Nk cells Perforin

Corresponding Organization : Karolinska Institutet

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Variable analysis

independent variables

Treatment with 50 nM CMA or DMSO

dependent variables

Perforin protein expression
CD107a protein expression

control variables

Time of treatment (3 hours)
Temperature (37°C)
Cell lysis procedure (as described in Fasbender et al., 2017)

controls

Positive control: Not specified
Negative control: DMSO treatment

Annotations

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