Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cytokine receptor signaling is required for the survival of ALK− anaplastic large cell lymphoma, even in the presence of JAK1/STAT3 mutations

doi: 10.1073/pnas.1700682114

Figure Lengend Snippet: JAK1 (and JAK2) are required for cell survival in JAK inhibitor-sensitive cells. (A) shJAK1 induced cell death in all JAK inhibitor-sensitive cells. The percentage of viable shRNA+ cells at days 2, 4, 6, 8, 10, and 12 after addition of doxycycline was compared with that of day 0. shRNA-infected cells were GFP+; the shRNA expression was induced by doxycycline. shSC4 served as a negative control; shRPL6, as a positive control. (B) shJAK2 induced cell death in Mac-1/2A/2B cells. (C) Western blot analysis of JAK1 and p-STAT3 in Mac-1 cells stably infected with retroviral shJAK1. The shRNA expression was induced by doxycycline for 24 h or 72 h. (D) Western blot analysis of PCM1-JAK2 and p-STAT3 in Mac-1 cells stably infected with retroviral shJAK2.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology: JAK1, JAK2 (3230), STAT3, and p-STAT3 (9145).

Techniques: shRNA, Infection, Expressing, Negative Control, Positive Control, Western Blot, Stable Transfection

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cytokine receptor signaling is required for the survival of ALK− anaplastic large cell lymphoma, even in the presence of JAK1/STAT3 mutations

doi: 10.1073/pnas.1700682114

Figure Lengend Snippet: JAK1 and STAT3 mutations induce increased STAT3 activity in response to IL-6. (A) JAK1G1097V induced increased STAT3 reporter activity compared with WT JAK1 in response to IL-6 (JAK1 vs. JAK1G1097V, P = 0.02). (B) JAK1G1097V induced greater STAT3 phosphorylation in response to IL-6. (C) STAT3 mutants induced increased STAT3 reporter activity in response to IL-6 compared with WT STAT3 (STAT3 vs. G618R, P < 0.01; STAT3 vs. S614R, P = 0.03; STAT3 vs. D661Y, P < 0.01). (D) STAT3 mutants induced increased STAT3 phosphorylation in response to IL-6. IL-6 (10 ng/mL) was added (or not) to the cell culture at 30 min before harvesting.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology: JAK1, JAK2 (3230), STAT3, and p-STAT3 (9145).

Techniques: Activity Assay, Cell Culture

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cytokine receptor signaling is required for the survival of ALK− anaplastic large cell lymphoma, even in the presence of JAK1/STAT3 mutations

doi: 10.1073/pnas.1700682114

Figure Lengend Snippet: WT JAK1 or STAT3 is capable of promoting cell growth in JAK1/STAT3 mutant-containing FE-PD cells. (A) WT JAK1 abrogated cell death induced by shJAK1. FE-PD cells that stably expressed WT were infected with shJAK1 to knock down endogenous JAK1 expression. Cell growth was monitored in infected cells (GFP+). shJAK1 expression was induced by doxycycline. (B) Western blot analysis of JAK1 expression in WT and JAK1-overexpressing cells. (C) WT STAT3 abrogated cell death induced by shSTAT3. (D) Western blot analysis of STAT3 expression in WT and STAT3-overexpressing cells.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology: JAK1, JAK2 (3230), STAT3, and p-STAT3 (9145).

Techniques: Mutagenesis, Stable Transfection, Infection, Expressing, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cytokine receptor signaling is required for the survival of ALK− anaplastic large cell lymphoma, even in the presence of JAK1/STAT3 mutations

doi: 10.1073/pnas.1700682114

Figure Lengend Snippet: Replacing endogenous JAK1 with WT JAK 1 did not affect pSTAT3 Y705 or pSTAT3 S727 expression. **Endogenous JAK1 was knocked down by shJAK1, whereas WT JAK1 was not knocked down.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology: JAK1, JAK2 (3230), STAT3, and p-STAT3 (9145).

Techniques: Expressing

Journal: International journal of oncology

Article Title: Macrophage-secreted IL-8 induces epithelial-mesenchymal transition in hepatocellular carcinoma cells by activating the JAK2/STAT3/Snail pathway.

doi: 10.3892/ijo.2014.2761

Figure Lengend Snippet: Figure 7. Role of IL-8 signaling in EMT. (A) The expression of Jak2, p-Jak2, Stat3, p-Stat3 and Snail were assessed in Hep-G2 exposure to different concentrations of IL-8. (B-D) Semi-quantitative analysis demonstrated that Jak2/Stat3/Snail pathway could be activated by IL-8. (#p>0.05; *p<0.05). (E) Immunofluorescent analysis of E-Cadherin, N-Cadherin and Snail in Hep-G2 vs. Hep-G2 that was incubated with IL-8. The green signal represents the staining of the corresponding protein and the blue signal represents the DAPI-stained nuclei. The morphological change of Hep-G2 after incubated with IL-8 was also observed. (F and G) Functional analysis is illustrated after IL-8 had been added to the media for 24 h. A transwell assay showed that the number of invading Hep-G2 cells were higher that in control groups (original magnification, x100).

Article Snippet: Rabbit anti-human JAK2/p-JAK2 monoclonal antibody (1:1,000; 3230; Cell Signaling Technology, Danvers, MA, uSA/1:1,000; 4406; Cell Signaling Technology) and rabbit anti-human STAT3/p-STAT3 monoclonal antibody (1:1,000; 4904; Cell Signaling Technology/1:2,000; 9145; Cell Signaling Technology) were selected to detect the JAK2/STAT3 pathway.

Techniques: Expressing, Incubation, Staining, Functional Assay, Transwell Assay, Control