Anti-ACK1 (phospho Y284) 抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACK1 (phospho Y284) antibody (AB74091)

ab74091 at 1/50 dilution staining ACK1 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue, in the absence or presence of the immunising peptide.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ACK1 (phospho Y284) antibody (AB74091)

ab74091 at 1/500 dilution staining ACK1 in A549 cells by Immunofluorescence, in the absence or presence of the immunising peptide.

• WB

Unknown

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

All lanes:

Western blot - Anti-ACK1 (phospho Y284) antibody (ab74091) at 1/500 dilution

Lane 1:

HepG2 cell extractstreated with EGF (200ng/ml, 30mins) at 30 µg

Lane 2:

HepG2 cell extractstreated with EGF (200ng/ml, 30mins) at 30 µg with immunising peptide

Predicted band size: 115 kDa

Observed band size: >117 kDa

false

• WB

CiteAb

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

Western Blotting using Anti-ACK1 (phospho Y284) antibody, ab74091. Publication image from Zhang, T. et al., 2020, Mol Cancer, 32404161. Legend direct from paper.

Activation of antiapoptotic signaling through the Ack1/AKT pathway contributes to ASK120067 resistance. a The levels of AKT phosphorylation (p-AKT) in NCI-H1975 and 67R cells were determined by immunoblotting analysis. b The inhibitory activity of ASK120067 on p-AKT expression in NCI-H1975 cells and 67R cells was compared. c Knockdown of Ack1 expression using short hairpin RNA (shRNA) decreased the levels of phosphorylated AKT in 67R cells. d The mRNA and protein levels of proapoptotic protein BIM in NCI-H1975 and 67R cells were determined by real-time PCR (left panel) and Western blot analysis (right panel), respectively. e The effect of ASK120067 on BIM expression in NCI-H1975 and 67R cells was examined. f Knockdown of Ack1 expression in 67R cells increased the expression of BIM by decreasing the phosphorylation of AKT. g to i, the combination of ASK120067 with Ack1 inhibitors synergistically suppressed AKT activation g and induced the transcription h and protein expression of BIM i

false

• WB

CiteAb

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

Western Blotting using Anti-ACK1 (phospho Y284) antibody, ab74091. Publication image from Zhang, T. et al., 2020, Mol Cancer, 32404161. Legend direct from paper.

Activation of Ack1 is sufficient to cause resistance to ASK120067, and drug combinations suppress proliferation and induce apoptosis of ASK120067-resistant cells. a Immunoblot analysis of total Ack1 and phosphorylated Ack1 (p-Ack1) levels in parental NCI-H1975 and 67R cells. b The expression of p-Ack1 in NCI-H1975 xenograft tumors and 67R xenograft tumors was compared by immunoblotting. c The p-Ack1 protein in AZDR and NCI-H1975 was detected by immunoblotting. d Correlation analysis of Ack1 expression and relapse-free survival (RFS) of 204 lung adenocarcinoma cancer patients (GSE22138) is presented as a Kaplan-Meier plot. e The antiproliferation effects of ASK120067 on NCI-H1975 cells ectopically expressing negative control vector or Ack1 were assessed using an SRB assay. f Knockdown of Ack1 expression in 67R effectively enhanced the antiproliferation potency of ASK120067. g ASK120067 in combination with AIM-100 caused a significantly higher growth-inhibition rate in ASK120067-resistant cells than that with ASK120067 treatment alone. h AIM-100 synergistically enhanced the apoptosis-inducing activity of ASK120067 in 67R cells. Cells were treated with ASK120067, AIM-100 alone or both drugs in combination for 48 h, and apoptosis was assessed using flow cytometry. i and j Combination ASK120067 with dasatinib i or bosutinib j partially restored the growth-inhibition sensitivity of ASK120067-resistant cells to ASK120067 treatment. k Combination ASK120067 with AIM-100, dasatinib or bosutinib synergistically inhibited the growth of AZDR cells. Data are plotted as the mean ± SD, and significance of differences was evaluated by Student’s t test (∗p < 0.05, ∗∗p < 0.01)

false

• WB

CiteAb

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

Western Blotting using Anti-ACK1 (phospho Y284) antibody, ab74091. Publication image from Zhang, T. et al., 2020, Mol Cancer, 32404161. Legend direct from paper.

Activation of Ack1 is sufficient to cause resistance to ASK120067, and drug combinations suppress proliferation and induce apoptosis of ASK120067-resistant cells. a Immunoblot analysis of total Ack1 and phosphorylated Ack1 (p-Ack1) levels in parental NCI-H1975 and 67R cells. b The expression of p-Ack1 in NCI-H1975 xenograft tumors and 67R xenograft tumors was compared by immunoblotting. c The p-Ack1 protein in AZDR and NCI-H1975 was detected by immunoblotting. d Correlation analysis of Ack1 expression and relapse-free survival (RFS) of 204 lung adenocarcinoma cancer patients (GSE22138) is presented as a Kaplan-Meier plot. e The antiproliferation effects of ASK120067 on NCI-H1975 cells ectopically expressing negative control vector or Ack1 were assessed using an SRB assay. f Knockdown of Ack1 expression in 67R effectively enhanced the antiproliferation potency of ASK120067. g ASK120067 in combination with AIM-100 caused a significantly higher growth-inhibition rate in ASK120067-resistant cells than that with ASK120067 treatment alone. h AIM-100 synergistically enhanced the apoptosis-inducing activity of ASK120067 in 67R cells. Cells were treated with ASK120067, AIM-100 alone or both drugs in combination for 48 h, and apoptosis was assessed using flow cytometry. i and j Combination ASK120067 with dasatinib i or bosutinib j partially restored the growth-inhibition sensitivity of ASK120067-resistant cells to ASK120067 treatment. k Combination ASK120067 with AIM-100, dasatinib or bosutinib synergistically inhibited the growth of AZDR cells. Data are plotted as the mean ± SD, and significance of differences was evaluated by Student’s t test (∗p < 0.05, ∗∗p < 0.01)

false

• WB

CiteAb

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

Western Blotting using Anti-ACK1 (phospho Y284) antibody, ab74091. Publication image from Zhang, T. et al., 2020, Mol Cancer, 32404161. Legend direct from paper.

Activation of Ack1 is sufficient to cause resistance to ASK120067, and drug combinations suppress proliferation and induce apoptosis of ASK120067-resistant cells. a Immunoblot analysis of total Ack1 and phosphorylated Ack1 (p-Ack1) levels in parental NCI-H1975 and 67R cells. b The expression of p-Ack1 in NCI-H1975 xenograft tumors and 67R xenograft tumors was compared by immunoblotting. c The p-Ack1 protein in AZDR and NCI-H1975 was detected by immunoblotting. d Correlation analysis of Ack1 expression and relapse-free survival (RFS) of 204 lung adenocarcinoma cancer patients (GSE22138) is presented as a Kaplan-Meier plot. e The antiproliferation effects of ASK120067 on NCI-H1975 cells ectopically expressing negative control vector or Ack1 were assessed using an SRB assay. f Knockdown of Ack1 expression in 67R effectively enhanced the antiproliferation potency of ASK120067. g ASK120067 in combination with AIM-100 caused a significantly higher growth-inhibition rate in ASK120067-resistant cells than that with ASK120067 treatment alone. h AIM-100 synergistically enhanced the apoptosis-inducing activity of ASK120067 in 67R cells. Cells were treated with ASK120067, AIM-100 alone or both drugs in combination for 48 h, and apoptosis was assessed using flow cytometry. i and j Combination ASK120067 with dasatinib i or bosutinib j partially restored the growth-inhibition sensitivity of ASK120067-resistant cells to ASK120067 treatment. k Combination ASK120067 with AIM-100, dasatinib or bosutinib synergistically inhibited the growth of AZDR cells. Data are plotted as the mean ± SD, and significance of differences was evaluated by Student’s t test (∗p < 0.05, ∗∗p < 0.01)

false

• WB

CiteAb

Western blot - Anti-ACK1 (phospho Y284) antibody (AB74091)

Western Blotting using Anti-ACK1 (phospho Y284) antibody, ab74091. Publication image from Zhang, T. et al., 2020, Mol Cancer, 32404161. Legend direct from paper.

Combination therapy with ASK120067 and Ack1 inhibitors showed synergistic in vivo antitumor effects in the 67R xenograft model. a Growth of 67R xenograft tumors following daily treatment with 5 mg/kg ASK120067, 25 mg/kg dasatinib or a combination of ASK120067 and dasatinib for 21 days. b The expression levels of p-Ack1 and p-AKT in 67R xenograft tumors were assessed after 21 days of treatment. c Cell apoptosis in 67R xenograft tumors was tested by the TUNEL assay after 21 days of treatment. Significance of differences was determined by Student’s t test (∗p < 0.05, ∗∗p < 0.01). d Proposed mechanism for the efficacy of the combination strategy in ASK120067-resistant NCI-H1975 cells

false