anti alex3  (Proteintech)

Journal: bioRxiv

Article Title: Brucella effector hijacks endoplasmic reticulum quality control machinery to prevent premature egress

doi: 10.1101/699330

Figure Lengend Snippet: (A) Co-immunoprecipitation (co-IP) from cell extracts expressing either HA-BspL, HA-BspB and HA-VceC using HA-trapping beads. Flow through and elutions were probed with antibodies against Alex3, Ubiquilin (Ubqln) and Herp in succession. The level of each effector bound to the beads was revealed with an anti-HA antibody and 15% of the input used for the co-IP shown (at the bottom). Molecular weights are indicated (KDa). (B) Representative confocal micrograph of HeLa cells expressing HA-BspL (green) and labelled for Herp (red). Scale bar corresponds to 5 µm.

Article Snippet: For western blotting the following antibodies were used: rabbit anti-FLAG (Sigma, #F7425) at 1/1000; rabbit anti-Alex3 (Sigma, # HPA000967) at 1/100; rabbit anti-Ubiquilin 2 (Abcam, #ab217056) at 1/1000; rabbit anti-Herp EPR9649 (Abcam, # ab150424) at 1/1000; mouse anti-HA (Covance, clone 16B12, ref. MMS-101R) at 1/1000; rabbit anti-TCR clone 3A8 (Invitrogen, #TCR1145) at 1/1000; mouse anti-myc antibody clone 9E10 at 1/1000; mouse anti-actin AC-40 (Sigma, #A4700) at 1/1000.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Expressing

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

doi: 10.1177/1010428317701441

Figure Lengend Snippet: Figure 1. The expression of Alex3 in lung cancer specimens. (a) Alex3 revealed strong cytosolic expression in normal bronchial epithelium cells; however, Alex3 showed negative or dim expression in (b) squamous cell carcinoma and (c) adenocarcinoma (magnification: 200×). (d) Alex was highly expressed in non-cancerous tissues than lung carcinoma (magnification: 200×). (e) Kaplan–Meier analysis results indicated that the overall survival of patients with positive Alex3 expression was significantly longer than those without.

Article Snippet: The sections were incubated with anti-Alex3 mouse monoclonal antibody (sc393752; 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for overnight, followed by biotinylated goat anti-mouse IgG secondary antibody.

Techniques: Expressing

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

doi: 10.1177/1010428317701441

Figure Lengend Snippet: Figure 2. The expression of Alex3 in lung cancer cell lines. The (a) protein and (b) mRNA levels of Alex3 was significantly higher in normal bronchial epithelium cell (HBE) than that in lung cancer cells.

Article Snippet: The sections were incubated with anti-Alex3 mouse monoclonal antibody (sc393752; 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for overnight, followed by biotinylated goat anti-mouse IgG secondary antibody.

Techniques: Expressing

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

doi: 10.1177/1010428317701441

Figure Lengend Snippet: Figure 3. Overexpression of Alex3 reduced lung cancer cell invasion and migration. (a) Western blot and (b) qPCR analysis of Alex3 protein and mRNA levels after overexpression or silencing of Alex3 in H1299 and A549 cells, respectively. (c) Sequent transwell and (d) wound-healing assay results suggested that overexpression or depletion of Alex3 suppressed or enhanced the invasive and migration abilities of H1299 or A549 cells. *p < 0.05; **p < 0.01.

Article Snippet: The sections were incubated with anti-Alex3 mouse monoclonal antibody (sc393752; 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for overnight, followed by biotinylated goat anti-mouse IgG secondary antibody.

Techniques: Over Expression, Migration, Western Blot, Wound Healing Assay

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

doi: 10.1177/1010428317701441

Figure Lengend Snippet: Figure 4. Overexpression of Alex3 downregulated Slug and upregulated E-cadherin. (a) When we overexpressed Alex3 in H1299 cells, WB results indicated that the expression of Slug was decreased and E-cadherin was increased. Correspondingly, Slug was upregulated and E-cadherin was downregulated by depleting Alex3 using siRNA. However, the other EMT-related proteins revealed no visible changes. (b) IF results suggested that membranous expression of E-cadherin was increased or decreased followed by upregulating/downregulating Alex3 in H1299 or A549 cells. However, the expression and sublocalization of occludin and ZO-1 revealed no visible changes.

Article Snippet: The sections were incubated with anti-Alex3 mouse monoclonal antibody (sc393752; 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for overnight, followed by biotinylated goat anti-mouse IgG secondary antibody.

Techniques: Over Expression, Expressing

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

doi: 10.1177/1010428317701441

Figure Lengend Snippet: Figure 5. Overexpression of Alex3 promoted the phosphorylation of AKT. (a) The phosphorylated AKT was decreased or increased followed by overexpressing or interfering Alex3 in H1299 or A549 cells. However, the other key signaling pathway proteins showed no obvious alteration. (b) Upregulation of p-AKT and Slug and downregulation induced by overexpression of Alex3 were reversed by adding AKT-specific inhibitor LY294002.

Article Snippet: The sections were incubated with anti-Alex3 mouse monoclonal antibody (sc393752; 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C for overnight, followed by biotinylated goat anti-mouse IgG secondary antibody.

Techniques: Over Expression, Phospho-proteomics