Fig 2: Smooth muscle contraction and CAS phosphorylation, but not myosin light chain phosphorylation, are regulated by c-Abl. a & b Contractile force was reduced in tissues from c-Ablsmko than in c-Abl-lox mice, which is time- and dose-dependent (n = 5–7). Contractile force is normalized to the force induced by 10-4 M ACh. c Immunoblot analysis was used to assess myosin light chain phosphorylation at Ser-19 of extracts from tracheal rings of c-Abl-lox and c-Ablsmko mice. Data are mean ± SE (n = 4). d ACh-induced CAS phosphorylation at Tyr-410 was reduced in tracheal rings from c-Ablsmko mice as compared to c-Abl-lox mice. Data are mean ± SE (n = 6). **P < 0.01. NS, not significant

Fig 3: Downstream targets of Src in mouse and cellular models of SBMA. a–c Immunoblots showing the levels of phosphorylated and total p130Cas, Stat3, p38MAPK, and Akt proteins in the spinal cord and skeletal muscle of 9-week-old AR-97Q mice treated with or without A419259 (a). Quantitative analyses were performed using densitometry (b, c). d–f Immunoblots showing the levels of the phosphorylated and total Src, p130Cas, Stat3, p38MAPK, and Akt proteins in DHT-treated NSC34 and C2C12 cells stably expressing AR-97Q with or without 0.2 µM of A419259 administration (d). Quantitative analyses were performed using densitometry (e, f). g–i Immunoblots showing the levels of the phosphorylated and total Src, p130Cas, Stat3, and p38MAPK proteins in the NSC34 and C2C12 cells transfected with mock or Src plasmids (g). Quantitative analyses were performed using densitometry (h, i). Quantitative analyses were performed on samples from n = 3 animals per group. Error bars indicate the s.e.m. Statistical analyses were performed using the unpaired two-sided t-test (b, c, e, f, h, i). *p < 0.05 and **p < 0.01. N.S., not significant. Source data are provided as a Source Data file (a–i)

Fig 4: Unaltered signaling in skin tumors of ACK1 ko mice. Western blot analysis of lysates skin tumors and tumor-free skin of DMBA/TPA-treated control and ACK1 ko mice for indicated proteins, with corresponding quantifications. Similar amount of proteins were applied per lane. Akt was used as loading control, since ß-actin was found to vary between tumors and tumor-free skin (n: 3–5/3–5; error bars are standard deviation; one-way ANOVA, *P = 0.05; **P = 0.01; ***P = 0.001; **** to P = 0.0001; The following primary antibodies were tested on the same blot membrane and share the loading control: (Ack1, keratin-14, pp130Cas), (pErk1/2, Erk1/2), (p130Cas, pAkt, and Akt).

Fig 5: p130Cas/BCAR1 and EREG expression are negatively correlated in CRC cell lines and in right-sided and earlier stages of colorectal adenocarcinomas. (A,F–H) BCAR1 and EREG mRNA levels in patient samples of the TCGA CoAd data set were analyzed. Patient numbers for each investigated subgroup are presented in Table S3A. (A,F) BCAR1/EREG expression correlation calculated with Spearman correlation in the indicated subgroups. Spearman coefficients (rs) are shown. (B) Cells were transfected with control or p130Cas siRNA and subsequently incubated with or without recombinant EREG (12.5 ng/mL) or vehicle control. Proliferation assays were performed as described in Materials and Methods. Statistical differences between growth curves were calculated with the CGGC permutation test. (C) Cells were transfected with control, p130Cas, or EREG siRNA. BCAR1 and EREG mRNA expression was quantified with RT qPCR. Shown are expression levels in relation to control siRNA (set to 1) from one representative out of three performed experiments. (D,E) Cells were treated with recombinant EREG (12.5 ng/mL) or vehicle control for 24 h. (D) BCAR1 mRNA expression was quantified with RT qPCR. (E) Expression and phosphorylation (p-p130Cas, pooled phospho-antibodies) in WCE (20 µg) was investigated by IB. Band intensities were quantified by densitometric analysis. The ratio of p-p130Cas and p130Cas was calculated and related to vehicle control (set to 1). (A–H) ns, not significant. * p < 0.05. ** p < 0.01. *** p < 0.001. si, short-interfering RNA.