Ab68194

For Western blot analysis, 10 μg of protein were loaded on NuPAGE Novex 4–12% bis-tris Gels (Invitrogen) and processed as described (32 (link)). The following antibodies were used: rabbit anti-FGFR1 (1:500; no. 9740, Cell Signaling Technology), goat anti-OSMRβ (1:500; AF662, R&D Systems), mouse anti-myotrophin (1:500; 611830, BD), rabbit anti-GAPDH (1:5000; no. 2118, Cell Signaling Technology), rabbit anti-pERK (1:500; no. 9101, Cell Signaling Technology), mouse anti-panERK (1:500; 612641, BD), rabbit anti-ACTN1 (1:1000; ab68194, Abcam), rabbit anti-ACTN4 (1:1000; ab108198, Abcam), mouse anti-αSMA (1:1000; A5228, Sigma-Aldrich), rabbit anti–hexokinase I (1:1000; no. 2024, Cell Signaling Technology), mouse anti-MAD2 (1:1000; 610679, BD), mouse anti-Myh (1:1000; LS-B6307, LSBio), mouse anti-myomesin (1:100; clone M5, gift of H. M. Eppenberger), mouse anti-PCNA (1:1000; 555566, Bionity), rabbit anti–phospho–Histone H3 (Ser¹⁰) (1:1000; no. 9701, Cell Signaling Technology), rabbit anti–phospho-MEK1/2 (1:1000; no. 3958, Cell Signaling Technology), mouse anti–Ral A (1:5000; 610222, BD), rabbit anti-RUNX1 (1:1000; ab92336, Abcam), rabbit anti-SDHA (1:1000; no. 5839, Cell Signaling Technology), mouse anti-TIMP1 (1:1000; MAB9801, R&D Systems), and anti-goat Alexa Fluor 594 (1:1000; A-21468, Invitrogen).

MIA PaCa-2 and CFPAC-1 human pancreatic cancer cell lines were obtained from the American Type Culture Collection. Dan-G human pancreatic cancer cells were provided by Daniel Billadeau (Mayo Clinic). The 6741 human patient–derived xenograft (PDX) cells were isolated from pancreatic tumors implanted in nude mice as described previously (Sagar et al., 2016 (link)) and were provided by the Mayo Clinic SPORE in Pancreatic Cancer. Tissues were collected from surgical resections for research purposes with written informed consent and were deidentified before use in research, and their use was approved by the Mayo Clinic Institutional Review Board (IRB). HPDE normal pancreas cells were obtained from Daniel Billadeau (Mayo Clinic). All cell lines were cultured in DMEM with 10% fetal bovine serum (FBS) and penicillin/streptomycin, except HPDE cells, which were maintained Keratinocyte-SFM with epidermal growth factor, bovine pituitary extract, and penicillin/streptomycin and regularly screened for mycoplasma by 4′,6-diamidino-2-phenylindole (DAPI) staining and PCR.
Antibodies used in this paper were α-actinin 1 (Santa-Cruz: sc1782 and Abcam: ab68194), α-actinin 4 (Abcam: ab108198), GAPDH (Cell Signaling: D16H11), Ki-67 (Cell Signaling: 12202), and p-ERK (Cell Signaling: 4377).

For western blot analysis, 10% of total IP lysate was used. Primary antibodies recognizing ACTN1 (ab68194), ACTN4 (ab108198), MYH9 (ab75590) from Abcam were used. After incubation with the rabbit secondary antibodies, protein signals were developed using HRP.