Image gauge version 3.1

Cells were collected in sodium dodecyl sulfate sample buffer. After sonication, cell lysates were subjected to electrophoresis on 5–20% polyacrylamide gels and transferred onto polyvinylidene difluoride membranes (ATTO). The membranes were probed with specific antibodies for poly(adenosine diphosphate-ribose) polymerase (PARP), JNK, c-Jun, phosphorylated-c-Jun (Ser63) (Cell Signaling Technology, Danvers, MA, USA) and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA). After washing, the membranes were incubated with secondary horse-radish peroxidase-conjugated antibodies. Signals were detected by means of enhanced chemiluminescence (GE Healthcare Japan, Tokyo, Japan) and scanned with an image analyzer LAS-4000 and Image Gauge (version 3.1) (Fuji Film, Tokyo, Japan). Band intensities were determined using ImageJ software [14 (link)].

Cells were harvested with sodium dodecyl sulfate sample buffer. After sonication, cell lysates were subjected to electrophoresis on 5–20% polyacrylamide gels and transferred onto polyvinylidene difluoride membranes (ATTO). Membranes were probed with specific antibodies for poly(adenosine diphosphate-ribose) polymerase (PARP), nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor, alpha (IκBα), phosphorylated IκBα (Ser32) (Cell Signaling Technology, Danvers, MA, USA) and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA). After washing, membranes were incubated with secondary horse-radish peroxidase-conjugated antibodies. Signals were detected by means of enhanced chemiluminescence (GE Healthcare Japan, Tokyo, Japan) and scanned with an image analyzer LAS-4000 and Image Gauge (version 3.1) (Fuji Film, Tokyo, Japan). Band intensities were determined by ImageJ software [26 (link)].

Cell lysates were collected in sodium dodecyl sulfate (SDS) sample buffer. After sonication, samples were subjected to electrophoresis on 5–20% polyacrylamide gels and transferred to polyvinylidene difluoride membranes (ATTO, Tokyo, Japan). The membranes were probed with specific antibodies to PACT, phosphorylation of PKR (Thr451), PKR (Santa Cruz Biotechnology), phosphorylation of IκBα (Ser32), IκBα (Cell Signaling Technology, Boston, MA) or GAPDH (Santa Cruz Biotechnology) for internal control. After a washing step, the membranes were incubated with secondary HRP-conjugated antibodies. Signals were detected with an ECL (GE Healthcare Japan, Tokyo, Japan) and scanned using an LAS-4000 image analyzer and Image Gauge (version 3.1) (Fuji Film, Tokyo, Japan) [6 (link)].