Journal: Journal of cellular physiology

Article Title: Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca 2+ ] and secretion in conjunctival goblet cells

doi: 10.1002/jcp.29846

Figure Lengend Snippet: Inhibition of protein kinase C (PKC) or calcium calmodulin-dependent kinase II (CaMK) blocks maresin 1 (MaR1)-stimulated increase in [Ca2+]i and high molecular weight glycoprotein secretion. Rat conjunctival goblet cells were incubated with the PKC inhibitor RO-317549 (10−7 M) or the CaMKII inhibitor KN93 (10−7 M) and its inactive control KN92 (10−7 M) for 30 min, then stimulated with MaR1 at 10−8 M for [Ca2+]i or 10−9 M for secretion or carbachol (Cch, 10−4 M). The change in peak [Ca2+]i (a and c) and secretion (b and d) are shown. Data are mean ± SEM from four (a), three (b), five (c), and three (d) experiments. *Significance above basal. #Significance from maresin 1 alone. [Ca2+]i, intracellular Ca2+; SEM, standard error of the mean

Article Snippet: VIP, U73122, {"type":"entrez-nucleotide","attrs":{"text":"U73343","term_id":"1688125","term_text":"U73343"}} U73343 , KN92, and KN93 were purchased from Tocris Bioscience (Ellisville, MO).

Techniques: Inhibition, High Molecular Weight, Incubation, Control

Journal: Nature Communications

Article Title: CaMKII induces permeability transition through Drp1 phosphorylation during chronic β-AR stimulation

doi: 10.1038/ncomms13189

Figure Lengend Snippet: ( a ) Pretreatment with β1-AR specific antagonist (CGP, 0.5 μM), but not β2-AR specific antagonist (ICI, 0.5 μM), blocked the ISO-induced flashes. N =38, 48, 36 and 31 cells from 3 to 5 rats in the groups of Vehicle, ISO, CGP+ISO and ICI+ISO, respectively. ( b ) Either the peptide inhibitor for PKA (PKI, 10 μM) or a cell-permeable inactive cAMP analogue (8-RP-cAMPs, 100 μM) failed to prevent the increase of flash after ISO treatment. N =25, 34, 20 and 19 cells from three rats in the groups of Vehicle, ISO, PKI+ISO and 8-RP+ISO, respectively. ( c ) CaMKII blockers, AIP (10 μM) and KN-93 (0.5 μM), inhibited ISO-induced flashes. KN-92, the inactive analogue of KN-93, was used as control. N =44, 55, 29, 30 and 31 cells from 3 to 5 rats in the groups of Vehicle, ISO, AIP+ISO, KN93+ISO and KN92+ISO, respectively. All the reagents in a – c were added to the myocytes 30 min before ISO treatment. ( d ) Inhibition of CaMKII activity by adenovirus-mediated overexpression of a dominant negative CaMKII (CaMKII DN) significantly attenuated mPTP openings 12 h after ISO treatment. N =20, 22, 18 and 20 cells from three rats in the groups of Vehicle, CaMKII DN, ISO and CaMKII DN+ISO, respectively. ( e ) CaMKII DN overexpression prolonged the time of laser-induced permanent loss of Δ ψ m after ISO treatment (100 nM, 48 h). N =208, 179, 260 and 165 mitochondria from 15 to 25 cells and three rats in the groups of Vehicle, CaMKII DN, ISO and CaMKII DN+ISO, respectively. ( f ) CaMKII DN overexpression prevented ISO-induced cell death (1 μM, 48 h). N =474, 577 and 374 cells from four rats in the group of Vehicle, ISO and CaMKII DN+ISO, respectively. Data in a – f are mean±s.e.m. * P <0.05 versus Control group, # P <0.05 versus ISO group. The data were analysed using One-way ANOVA followed by Turkey post test in a – f .

Article Snippet: KN93 and KN92 were purchased from Merck Millipore Corporation (La Jolla, CA, USA).

Techniques: Control, Inhibition, Activity Assay, Over Expression, Dominant Negative Mutation