shrna against lif  (Santa Cruz Biotechnology)

Journal: Immunity, Inflammation and Disease

Article Title: Modified Hongteng Baijiang decoction enema improves sequelae of pelvic inflammatory disease by regulating the LIF/JAK2/STAT3 pathway and gut microbiota

doi: 10.1002/iid3.1300

Figure Lengend Snippet: Effect of MHTBD enemas on the endometrial LIF/JAK2/STAT3 signaling pathway. Relative protein expression of LIF, JAK2, p‐JAK2, STAT3, and p‐STAT3 in endometrial tissue ( n = 3); protein bands were calculated as a ratio relative to β‐actin protein levels. The data are expressed as the mean ± SD. Compared with the Sham group, ** p < .01; compared with the SPID group, # p < .05, ## p < .01; ns p > .05. MHTBD, modified Hongteng Baijiang decoction; SPID, sequelae of pelvic inflammatory disease.

Article Snippet: Electrophoresis was performed on 12% sodium dodecyl sulfate‒polyacrylamide gels and transferred to PVDF membranes, which were blocked with 5% BSA in TBST for 60 min. Primary antibodies against LIF (cat. no. A1288, 1:1000), JAK2 (cat. no. A7694, 1:1000), p‐JAK2 (cat. no. AP0531, 1:1000), STAT3 (cat. no. A1192, 1:1000), p‐STAT3 (cat. no. AP0715, 1:1000), and the internal reference β‐actin (cat. no. AC004, 1:2000) (ABclonal) were added, and the membranes were incubated overnight at 4°C.

Techniques: Expressing, Modification

Journal: Immunity, Inflammation and Disease

Article Title: Modified Hongteng Baijiang decoction enema improves sequelae of pelvic inflammatory disease by regulating the LIF/JAK2/STAT3 pathway and gut microbiota

doi: 10.1002/iid3.1300

Figure Lengend Snippet: Effect of the MHTBD‐mediated LIF/JAK2/STAT3 signaling pathway on endometrial pathology and receptivity in rats. Endometrial tissue was observed via H&E staining (magnification, ×100 and ×400; scale bars, 200 and 50 μm; n = 3) (A). Scanning electron microscopy (magnification, ×5000; scale bar, 5 μm) (B). Immunohistochemical staining of ER, PR, ITGB3, and CD31 in endometrial tissue (magnification, ×40; scale bar, 50 μm; n = 3) (C). Number of blastocysts ( n = 12) (D). Positive expression of ER, PR, ITGB3, and CD31 in endometrial tissue ( n = 3) (E). Relative protein expression of JAK2, p‐JAK2, STAT3, and p‐STAT3 in endometrial tissue ( n = 3). The protein band density was calculated as the ratio relative to the β‐actin protein concentration (F). The data are expressed as the mean ± SD. Compared with the Sham group, ** p < .01; compared with the SPID group, # p < .05, ## p < .01; compared with the SPID + EC330 group, ΔΔ p < .01; compared with the SPID + MHTBD group, ▲ p < .05, ▲▲ p < .01; ns p > .05. ER, estrogen receptor; H&E, hematoxylin−eosin; ITGB3, integrin β3; MHTBD, modified Hongteng Baijiang decoction; SPID, sequelae of pelvic inflammatory disease.

Article Snippet: Electrophoresis was performed on 12% sodium dodecyl sulfate‒polyacrylamide gels and transferred to PVDF membranes, which were blocked with 5% BSA in TBST for 60 min. Primary antibodies against LIF (cat. no. A1288, 1:1000), JAK2 (cat. no. A7694, 1:1000), p‐JAK2 (cat. no. AP0531, 1:1000), STAT3 (cat. no. A1192, 1:1000), p‐STAT3 (cat. no. AP0715, 1:1000), and the internal reference β‐actin (cat. no. AC004, 1:2000) (ABclonal) were added, and the membranes were incubated overnight at 4°C.

Techniques: Staining, Electron Microscopy, Immunohistochemical staining, Expressing, Protein Concentration, Modification

Journal: Cell Death & Disease

Article Title: Leukemia inhibitory factor drives glucose metabolic reprogramming to promote breast tumorigenesis

doi: 10.1038/s41419-022-04820-x

Figure Lengend Snippet: A Ectopic LIF expression (left panel) as well as treatment with recombinant human LIF protein (rhLIF, 100 ng/ml for 6 h; right panel enhanced glucose uptake in breast cancer cell lines, including MCF7, MDA-MB 231, T47D, MDA-MB 468, ZR-75-1 and SKBR3 cells as determined by measuring the uptake of 3 H-2-DG in cells. The overexpression of LIF in these cells was shown in Supplementary Fig . B Knockdown of endogenous LIF by two different shRNAs reduced glucose uptake in MCF7 and MDA-MB 231 cells. The knockdown of LIF expression in these cells was shown in Supplementary Fig . C Ectopic LIF expression increased glucose uptake in the xenograft tumors formed by MDF7 and MDA-MB 231 cells as determined by measuring the uptake of 3 H-2-DG in tumor tissues. D . High LIF expression was associated with the increased 18 F-FDG uptake in human breast tumors. Left panels: representative PET scan data and images of LIF IHC staining. SUV: standardized uptake value. In A – C , n = 3/group; in D , n = 18 for total patient samples. *: p < 0.05; **: p < 0.01; ***: p < 0.001; unpaired Student’s t -test for A – C ; correlation in D was calculated by Spearman’s Rho correlation analysis.

Article Snippet: Antibody against LIF (anti-LIF; 39N7D10, Novus, 1:500) was used for IHC staining.

Techniques: Expressing, Recombinant, Over Expression, Knockdown, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Leukemia inhibitory factor drives glucose metabolic reprogramming to promote breast tumorigenesis

doi: 10.1038/s41419-022-04820-x

Figure Lengend Snippet: A Glut1 mRNA levels were detected by quantitative real-time PCR (qPCR) in MCF7, MDA-MB 231, and T47D cells with or without ectopic LIF expression. B , C Ectopic LIF expression ( B ) or rhLIF treatment (100 ng/ml for 12 h) ( C ) promoted endogenous Glut1 PM translocation in MCF7, MDA-MB 231, and T47D cells as determined by Western-blot assays. D Knockdown of LIF decreased endogenous Glut1 PM translocation in MDA-MB 231 cells. E Ectopic LIF expression increased the PM translocation of ectopically expressed Myc-Glut1 in MCF7, MDA-MB 231 and T47D cells as determined by Western-blot assays. F LIF neutralization antibody (LIF neu-ab) largely abolished exogenous Glut1 PM translocation promoted by LIF. G The rhLIF treatment promoted exogenous Glut1 PM translocation in cells. H Knockdown of LIF decreased Myc-Glut1 PM translocation in MDA-MB 231 cells. I Ectopic LIF expression promoted Myc-Glut1 PM translocation (left panels) while knockdown of LIF decreased Myc-Glut1 PM translocation (right panels) in MDA-MB 231 cells as determined by IF staining assays. Scale bar, 10 μm. J Ectopic LIF expression promoted the PM translocation of Myc-Glut1 in MCF7, MDA-MB 231, and T47D cells as determined by flow cytometry assays. Left panels: representative images of flow cytometry analysis. Right panels: quantifications of relative fluorescence intensity of Myc-Glut1 on the cell membrane normalized with total Myc-Glut1 fluorescence intensity in cells. In A , J data are presented as mean ± SD. n = 3/group. * p < 0.05; ** p < 0.01; NS: non-significant; unpaired Student’s t -test. Uncropped Wes t ern-blot images are shown in Supplementary Fig .

Article Snippet: Antibody against LIF (anti-LIF; 39N7D10, Novus, 1:500) was used for IHC staining.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Translocation Assay, Western Blot, Knockdown, Neutralization, Staining, Flow Cytometry, Fluorescence, Membrane