rabbit  (Novus Biologicals)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Selective targeting of α7 nicotinic acetylcholine receptor by synthetic peptide mimicking loop I of human SLURP-1 provides efficient and prolonged therapy of epidermoid carcinoma in vivo

doi: 10.3389/fcell.2023.1256716

Figure Lengend Snippet: (A) Real-time PCR analysis of the mRNA expression of genes coding α7-nAChR ( CHRNA7 ), PTEN ( PTEN ), integrin α5 ( ITGA5 ), ceramide kinase ( CERK ), SLURP-1 ( SLURP1 ), KLF4 ( KLF4 ) in mice xenografted tumors after the 10-day treatment with SLURP-1 (0.5 mg/kg), Oncotag (0.125 mg/kg), or Oncotag (1.25 mg/kg) and 11 subsequent days of rest. Data are presented as lg of the mRNA expression level normalized to the expression of the same gene in the control group (mice treated with saline, 0, dashed line) ± SEM (n = 7–9). For each sample the gene expression was pre-normalized to expression of ACTB , GAPDH , and RPL13A genes of housekeeping proteins. * ( p < 0.05), ** ( p < 0.01), and *** ( p < 0.001) indicate significant differences from control group (0, level), by a two-tailed one-sample t -test, followed by the Holm-Sidak’s post hoc test. (B) Representative Western blot membrane with analysis of the KLF4 expression in tumors after the saline (control), SLURP-1, and Oncotag (0.125 mg/kg) treatment. Whole membranes are in . (C) KLF4 expression on a protein level normalized to control (saline, 1, dashed line) group ± SEM (n = 4). The KLF4 level on each blot was pre-normalized to the total protein level according to ponceau S staining . * ( p < 0.05) and *** ( p < 0.001) indicate significant difference from the control (1, level) by one-sample two-tailed t -test followed by Holm-Sidak’s post hoc test.

Article Snippet: Western blotting was performed with primary anti-KLF4 antibodies (NBP2-24749, Novus Bio, Centennial, United States, 1:2000) and secondary anti-rabbit antibodies (111-035-003, Jackson Immunoresearch, 1:5000).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Control, Saline, Gene Expression, Two Tailed Test, Western Blot, Membrane, Staining

Journal: Scientific reports

Article Title: Smooth muscle cell specific NEMO deficiency inhibits atherosclerosis in ApoE -/- mice.

doi: 10.1038/s41598-022-16737-8

Figure Lengend Snippet: Figure 5. Deletion of NEMO in smooth muscle cells inhibits their phenotype switching. Representative pictures from immunostainings of atherosclerotic lesions of ApoE−/− or NEMOSMCiKO/ApoE−/− mice with antibodies against KLF4 (a) and α-SMA (c). Scale bars = 0.1 mm. Graph showing quantification of KLF4 positive cells (b) and α-SMA content (d) in the lesions of ApoE−/− (n = 13) or NEMOSMCiKO/ApoE−/− (n = 10) mice (Mann–Whitney test). (e) Cultured smooth muscle cells were stimulated with TNF (1 ng/ml) and LPS (10 ng/ ml) or left unstimulated in starved medium for 48 h. Total protein lysates of smooth muscle cells were subjected to western blot analysis of KLF4 and NEMO protein expression. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. Uncropped blots are presented in Supplementary Fig. 4.

Article Snippet: The immunoblots were incubated overnight with primary antibodies against KLF4 (NBP2-24749, Novus Biological, 1:200), NEMO (homemade rabbit polyclonal serum), and GAPDH (NB300-221, 1:5000).

Techniques: MANN-WHITNEY, Cell Culture, Western Blot, Expressing, Control