PKN2 Antibody (509105)

Detection of Human/Mouse/Rat PKN2 by Western Blot.

Western blot shows lysates of A431 human epithelial carcinoma cell line, TF-1 human erythroleukemic cell line, C2C12 mouse myoblast cell line, NIH-3T3 mouse embryonic fibroblast cell line, L6 rat myoblast cell line, and Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat PKN2 Monoclonal Antibody (Catalog # MAB5686) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for PKN2 at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of PKN2 by Western Blot

PKN2 modulates TEAD-driven transcription and nuclear localization of the mechanosensor YAP(A) Schematic showing potential downstream targets of PKN2 involved in myofibroblast differentiation.(B–D) Normalized expression of SRF (B), TEAD (C), or SMAD (D) responsive Firefly luciferase reporter in WT and KO PSCs starved in 0.5–1% serum or treated with 5 ng/mL TGF-beta 1 or 10% serum. Values are normalized to a Renilla luciferase control per sample and presented relative to WT serum-starved PSCs (n = 5; two-way ANOVA with Tukey's correction).(E) qPCR analysis of expression of indicated genes in PKN2 WT and KO PSCs expressed as a fold change to WT control (n = 4).(F) Immunofluorescent images of YAP1 localization (green) in WT and PKN2KO PSCs plated at low and high density on glass coverslips for 48 h (minimum of 100 cells/condition; n = 3; scale bar represents 50 μm).(G) Percentage of WT and PKN2KO PSCs with YAP-positive nuclei plated at both high and low density (n = 3; unpaired t test).(H) Quantification by Python CellProfiler algorithm of YAP nuclear intensity for indicated number of cell neighbors (n = 3; two-way ANOVA with Sidak's test).(I and J) Representative western blot and quantification of p-YAP S112 and total YAP expression in WT and PKN2KO PSCs plated at low and high density (n = 3; two-way ANOVA with Tukey's multiple comparisons test).(K and L) Western blot and quantification of p-SMAD2/3 induction with 5 ng/mL TGF-beta 1 for indicated time points; quantification expressed relative to untreated WT PSCs (n = 3; unpaired t test).(M) Western blot analysis of p-p70 S6K, total p70 S6K, p-ERK1/2, and total ERK in WT and PKN2KO PSCs starved in 1% serum and treated with vehicle or 5 ng/mL TGF-beta 1 for 4 h (n = 2).For statistics: ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; and ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35081338), licensed under a CC-BY license. Not internally tested by R&D Systems.