keratin 1 rabbit polyclonal customized peptide specific antibody  (Peptide Specialty Laboratories)

Journal: iScience

Article Title: A feedback loop between plakophilin 4 and YAP signaling regulates keratinocyte differentiation

doi: 10.1016/j.isci.2024.110762

Figure Lengend Snippet: PKP4 modulates keratinocyte differentiation (A) Protein levels of differentiation markers in WT and PKP4-KO cells grown for 24h or 72h in medium with or without Ca 2+ . Left: Representative western blots of epidermal differentiation markers. Ponceau S staining was used as a loading control. For filaggrin, the main band representing the filaggrin three-domain intermediate was quantified (marked with an arrow). Right: Quantification of protein amounts normalized to Ponceau S staining and relative to WT cells grown for 24h in medium without Ca 2+ . Averages +SD from five independent experiments are plotted. Quantification of LCM versus HCM treatment has been omitted. See also Figure S1 . (B) Immunofluorescence analysis of keratin 1 in WT and PKP4-KO cells grown for 24h in HCM. Left: Representative immunofluorescence images showing desmoplakin (DSP) and keratin 1 localization. Scale bar = 50 μm, detail 10 μm. Right: Mean keratin 1 fluorescence intensity. n ≥ 100 cells per condition from two independent experiments. (C) Confocal immunofluorescence analysis of keratin 1 in WT and PKP4-KO cells grown for 72h in HCM. Typical example of confocal microscopy z-stacks showing maximum intensity projection (left panel), 3D project x axis rotation (mid panel), and orthogonal views (right panel) of PKP4-KO cells versus WT cells. Scale bar = 100 μm. (D) Quantification of the amounts of overlapping nuclei in WT or PKP4-KO cells, as determined from confocal images. Averages +SD from three independent experiments are plotted. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, not significant. Significance was determined by one-way ANOVA with Tukey’s multiple comparisons test (A) or by student’s unpaired two-tailed t-test (B and D).

Article Snippet: Keratin 1 rabbit polyclonal customized peptide specific antibody , Peptide Specialty Laboratories , N/A.

Techniques: Western Blot, Staining, Control, Immunofluorescence, Fluorescence, Confocal Microscopy, Two Tailed Test

Journal: iScience

Article Title: A feedback loop between plakophilin 4 and YAP signaling regulates keratinocyte differentiation

doi: 10.1016/j.isci.2024.110762

Figure Lengend Snippet: PKP4 promotes YAP target gene expression and YAP affects proliferation and differentiation of keratinocytes (A) mRNA amounts of YAP/TAZ targets in PKP4-KO and PKP4-KO+PKP4 (Rescue) cells relative to WT cells grown for 24h in LCM (Top) or HCM (Bottom). Eif3k was used as a reference gene. Averages +SD from three independent experiments are plotted. (B) mRNA amounts of YAP/TAZ targets in siRNA-treated WT cells grown for 24h in LCM (Top) or HCM (Bottom). Eif3k was used as a reference gene. Averages +SD from three independent experiments are plotted. (C) Measurement of proliferation using an IncuCyte S3 system. Top: Representative live cell images of WT, PKP4-KO, and PKP4-KO+PKP4 (Rescue) cells treated with control, YAP- or TAZ directed siRNAs grown for up to 96h in LCM. Scale bar = 100 μm. Bottom: Area occupied by cell nuclei at the indicated time points, as determined from live cell images. Data are shown relative to measurements at the beginning of recording (time zero). Graphs represent averages ±SD from three independent experiments. See also Figure S3 A. (D) Western Blots showing proliferation and differentiation marker expression in siRNA-treated WT and PKP4-KO cells grown for 24h in medium with or without Ca 2+ . Top: Representative western blots of YAP, TAZ, the proliferation marker phospho-RB, and the differentiation marker keratin 1. GAPDH was used as a loading control. Bottom: Quantification of protein amounts normalized to GAPDH and relative to WT cells grown in medium without Ca 2+ . Quantification of LCM versus HCM treatment has been omitted. Averages +SD from three independent experiments are plotted. See also Figure S3 B. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, not significant. Significance was determined by one-way ANOVA with Tukey’s multiple comparisons test (A, B, C, and D).

Article Snippet: Keratin 1 rabbit polyclonal customized peptide specific antibody , Peptide Specialty Laboratories , N/A.

Techniques: Targeted Gene Expression, Control, Western Blot, Marker, Expressing

Journal: iScience

Article Title: A feedback loop between plakophilin 4 and YAP signaling regulates keratinocyte differentiation

doi: 10.1016/j.isci.2024.110762

Figure Lengend Snippet:

Article Snippet: Keratin 1 rabbit polyclonal customized peptide specific antibody , Peptide Specialty Laboratories , N/A.

Techniques: Transduction, Virus, Recombinant, Formulation, Random Hexamer Labeling, Reverse Transcription, Western Blot, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Expressing, Mutagenesis, Plasmid Preparation, Software