Journal: Cancer Management and Research

Article Title:

HOXA10 deteriorates gastric cancer through activating JAK1/STAT3 signaling pathway

doi: 10.2147/cmar.s201342

Figure Lengend Snippet: Figure 3 Over-expression of HOXA10 promoted the activation of JAK1/STAT3 signaling in gastric cancer. OE-HOXA10, OE-NC, sh-HOXA10 and sh-NC lentivirus vectors were transiently transfected into MKN28 and NCI-N87 cells, then (A–B) The expression of proteins in JAK family (JAK1-3 and Tyk2) and their protein phosphorylation levels were assessed by Western blotting assay. (C–D) Both total and phosphorylated levels (p-) of STAT3 and STAT5B were determined by Western blotting. (E) Immunofluorescence assay was used to decide the effects of HOXA10 expression on STAT3 subcellular location. (*P<0.05, **P<0.01, sh-HOXA10 group compared with sh- NC group; #P<0.05, ##P<0.01, OE-HOXA10 group compared with OE-NC group).

Article Snippet: Primary antibodies used in this experiment are as follows: HOXA10 (1:1,000 dilution; No. ab191470; Abcam), JAK1 (1:1,000 dilution; No. 3332, Cell Signaling Technology, CA, USA), p-JAK1 (1:1,000 dilution; No. 3331, Cell Signaling Technology), JAK2 (1:1,000 dilution; No. 3230, Cell Signaling Technology), p-JAK2 (1:1,000 dilution; No. 3771, Cell Signaling Technology), JAK3 (1:1,000 dilution; No. 3775, Cell Signaling Technology), p-JAK3 (1:1,000 dilution; No. 5031, Cell Signaling Technology), Tyk2 (1:1,000 dilution; No. 9312, Cell Signaling Technology), p-Tyk2 (1:1,000 dilution; No. 9321, Cell Signaling Technology), STAT3 (1:1,000 dilution; No. 9139, Cell Signaling Technology), p-STAT3 (1:1,000 dilution; No. 4113, Cell Signaling Technology), STAT5B (1:1,000 dilution; No. 34,662, Cell Signaling Technology), p-STAT5B (1:1,000 dilution; No. ab52211, Abcam) and GAPDH (1:5,000 dilution; No. TA-08, Zhongshanjinqiao).

Techniques: Over Expression, Activation Assay, Transfection, Expressing, Phospho-proteomics, Western Blot

Journal: Cancer Management and Research

Article Title:

HOXA10 deteriorates gastric cancer through activating JAK1/STAT3 signaling pathway

doi: 10.2147/cmar.s201342

Figure Lengend Snippet: Figure 4 HOXA10 promoted cell viability and inhibited cell apoptosis through activating JAK1 signaling in gastric cancer. (A–B) qPCR and Western blotting assays were executed to determine the mRNA and protein expression of JAK1 after MKN28 or NCI-N87 cells were transfected with sh-JAK1 or sh-NC. (C–D) MTT assay was used to test cell proliferation after MKN28 or NCI-N87 cells were transfected with OE-HOXA10 or OE-HOXA10+sh-JAK1. (E) Cell cloning formation abilities were assessed by cloning formation assay after MKN28 or NCI-N87 cells were stably transfected with OE-HOXA10 or OE-HOXA10+sh-JAK1. (F) Cell apoptosis population was determined by flow cytometry assay after MKN28 or NCI-N87 cells were transfected with OE-HOXA10 or OE-HOXA10+sh-JAK1. (*P<0.05, **P<0.01, OE-HOXA10 group compared with control group; #P<0.05, OE-HOXA10+sh-JAK1 group compared with OE-HOXA10 group).

Article Snippet: Primary antibodies used in this experiment are as follows: HOXA10 (1:1,000 dilution; No. ab191470; Abcam), JAK1 (1:1,000 dilution; No. 3332, Cell Signaling Technology, CA, USA), p-JAK1 (1:1,000 dilution; No. 3331, Cell Signaling Technology), JAK2 (1:1,000 dilution; No. 3230, Cell Signaling Technology), p-JAK2 (1:1,000 dilution; No. 3771, Cell Signaling Technology), JAK3 (1:1,000 dilution; No. 3775, Cell Signaling Technology), p-JAK3 (1:1,000 dilution; No. 5031, Cell Signaling Technology), Tyk2 (1:1,000 dilution; No. 9312, Cell Signaling Technology), p-Tyk2 (1:1,000 dilution; No. 9321, Cell Signaling Technology), STAT3 (1:1,000 dilution; No. 9139, Cell Signaling Technology), p-STAT3 (1:1,000 dilution; No. 4113, Cell Signaling Technology), STAT5B (1:1,000 dilution; No. 34,662, Cell Signaling Technology), p-STAT5B (1:1,000 dilution; No. ab52211, Abcam) and GAPDH (1:5,000 dilution; No. TA-08, Zhongshanjinqiao).

Techniques: Western Blot, Expressing, Transfection, MTT Assay, Cloning, Tube Formation Assay, Stable Transfection, Cytometry, Control

Journal: Cancer Management and Research

Article Title:

HOXA10 deteriorates gastric cancer through activating JAK1/STAT3 signaling pathway

doi: 10.2147/cmar.s201342

Figure Lengend Snippet: Figure 6 HOXA10 promoted NCI-N87 cell tumorigenesis through activating JAK1/STAT3 signaling. NCI-N87 cells were stably transfected with OE-HOXA10, OE-HOXA 10+sh-JAK1, OE-HOXA1-+sh-STAT3 or their negative control vectors; then, the stable cell lines (5×106 cells for each group) were injected into the nude mice. And all mice were killed 28 days post-implantation and the image showed the tumor shape and average weights of each group. (*P<0.05, OE-HOXA10 group compared with control group; #P<0.05, OE-HOXA10+sh-JAK1 group or OE-HOXA10+sh-STAT3 group compared with OE-HOXA10 group).

Article Snippet: Primary antibodies used in this experiment are as follows: HOXA10 (1:1,000 dilution; No. ab191470; Abcam), JAK1 (1:1,000 dilution; No. 3332, Cell Signaling Technology, CA, USA), p-JAK1 (1:1,000 dilution; No. 3331, Cell Signaling Technology), JAK2 (1:1,000 dilution; No. 3230, Cell Signaling Technology), p-JAK2 (1:1,000 dilution; No. 3771, Cell Signaling Technology), JAK3 (1:1,000 dilution; No. 3775, Cell Signaling Technology), p-JAK3 (1:1,000 dilution; No. 5031, Cell Signaling Technology), Tyk2 (1:1,000 dilution; No. 9312, Cell Signaling Technology), p-Tyk2 (1:1,000 dilution; No. 9321, Cell Signaling Technology), STAT3 (1:1,000 dilution; No. 9139, Cell Signaling Technology), p-STAT3 (1:1,000 dilution; No. 4113, Cell Signaling Technology), STAT5B (1:1,000 dilution; No. 34,662, Cell Signaling Technology), p-STAT5B (1:1,000 dilution; No. ab52211, Abcam) and GAPDH (1:5,000 dilution; No. TA-08, Zhongshanjinqiao).

Techniques: Stable Transfection, Transfection, Negative Control, Injection, Control

Journal: medRxiv

Article Title: Reduced risk of placental parasitaemia associated with complement fixation on Plasmodium falciparum by antibodies among pregnant women

doi: 10.1101/2021.06.02.21258254

Figure Lengend Snippet: (A) Complement C5-9 fixation on CS2 pRBCs was tested by flow cytometry following opsonization with either antibodies from a subset of 7 PNG donor samples (with moderate to high C1q/C3 fixing activity on CS2 pRBCs) or 6 Australian malaria non-exposed donors in the presence of NS. Complement fixation is presented as the average gMFI of each sample tested in duplicate and the red line represents the median gMFI for all samples tested in each category. (B) Correlation between complement C5-9 fixation and C1q/C3 fixation tested on a subset of 7 PNG donor samples (with moderate to high C1q/C3 fixing activity on CS2 pRBCs). (C) Complement-mediated pRBC killing/lysis was examined by culturing CS2 pRBCs opsonized with antibodies from PNG pregnant women (n=7) in the presence of normal serum, heat-inactivated serum or no-added serum and measuring parasitaemia by flow cytometry after two growth cycles. Data are presented as the average parasitaemia of each sample run in triplicate over 3 independent experiments. The dotted red line represents the median parasitaemia of CS2 pRBCs cultured in the absence of either PNG or Australian malaria non-exposed antibodies and NS or HIS. Comparisons were made using Mann-Whitney U-test.

Article Snippet: Pigmented trophozoite stage parasites at ∼10% parasitaemia and 0.5% haematocrit were incubated with 25% NS and 10% antibodies from PNG pregnant women or Australian malaria non-exposed donors for 1 hour at 37°C. pRBCs were then washed in 0.5% BSA in PBS and C1q, C3 and C5-9 detected by staining with polyclonal rabbit anti-human C1q, goat anti-human C3 or monoclonal mouse anti-human C5-9 antibodies followed by respective polyclonal Alexa-488 conjugated goat anti-rabbit, rabbit anti-goat or goat anti-mouse antibodies. pRBCs were detected by staining with ethidium bromide (Bio-Rad) at 1/1000 dilution.

Techniques: Flow Cytometry, Activity Assay, Lysis, Cell Culture, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Resveratrol Induces Cell Cycle Arrest and Apoptosis in Malignant NK Cells via JAK2/STAT3 Pathway Inhibition

doi: 10.1371/journal.pone.0055183

Figure Lengend Snippet: Cells were treated with 50 µM of resveratrol for 24, 36 and 48 h. 50 µg of whole cell extract was prepared for each cell line, separated on SDS-PAGE and subjected to Western blotting with antibodies specific to phosphorylated JAK1 (p-JAK1), JAK1, phosphorylated TYK2 (p-TYK2), phosphorylated Akt (p-Akt), phosphorylated PTEN (p-PTEN), phosphorylated JAK2 (p-JAK2) and JAK2. The blots were stripped and reprobed with anti-α-tubulin antibody to show equal protein loading. The figures shown are representative results of three independent experiments.

Article Snippet: Antibodies against survivin, myeloid leukaemia cell differentiation protein 1 (MCL1), p21 Waf1/Cip1, p53, cdc2, cdk2, Bcl-2, Bcl-10, cleaved caspase-3 (Asp175), phosphorylated STAT3 (Tyr705), acetylated STAT3 (Lys685), phosphorylated PTEN (Ser380/Thr382/383), phosphorylated tyrosine kinase 2 (TYK2, Tyr1054/1055), phosphorylated Akt (Thr308), phosphorylated JAK1 (Tyr1022/1023), total JAK1, and phosphorylated JAK2 (Tyr1007/1008) were acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques: SDS Page, Western Blot

Journal: PLoS ONE

Article Title: Interim report on the effective intraperitoneal therapy of insulin-dependent diabetes mellitus in pet dogs using “Neo-Islets,” aggregates of adipose stem and pancreatic islet cells (INAD 012-776)

doi: 10.1371/journal.pone.0218688

Figure Lengend Snippet: AntiASC and anti-IC IgG responses as assessed by FACS in sera of the study dogs before (A) and after (B) treatment with allogeneic cNIs. Shown are percentages of FITC-labeled anti-dog IgG antibody. Sera were collected from dogs before (A) and 1.5 to 3 months after (B) cNI administration. The percent of positive cells is indicated above each column. Dogs do not show increased IgG responses to either ASCs or ICs after cNI administration, indicating there is no additional allo-immune response by the recipients to either cell type. Three dogs show pre-existing antibodies to ICs prior to treatment with cNIs, suggesting they have an autoimmune form of diabetes. (C) Results of ELISA testing of sera from the treated dogs for specific anti-islet antigens, IA2 and GAD65, indicate that none of the dogs have antibodies to GAD65 antigen, but that WSU-01’s serum contained antibodies to IA2 prior to, but not after treatment. Samples were run in duplicate. Note: a pre-treatment serum sample was not available for VSH-01.

Article Snippet: ELISAs for GAD65 and IA2 autoantibodies were kindly carried out by Dr. Boris Fehse, University of Hamburg, Germany, using GAD65 and IA2 ELISA kits (both from Euroimmun, Lübeck, Germany, EA 1022–9601 G and EA 1023–9601 G, respectively) and the same serum samples were analyzed for allo-antibody levels.

Techniques: Labeling, Enzyme-linked Immunosorbent Assay