The Magazine "Student" and its Times (1965 - 1968)

Hlavním cílem této práce, kterou jsem nazval "Casopis Student a jeho doba", je postihnout význam casopisu v dejinách naší žurnalistiky a prostrednictvím obsahu clánku a osudu lidí podílejících se na jeho existenci priblížit atmosféru doby, v níž Student vycházel. Tato práce je svým zpusobem nová a zasahuje do mnoha oblastí kulturního i politického života druhé poloviny šedesátých let dvacátého století. Pri vytvárení koncepce obsahu práce jsem se tady nemohl oprít o žádnou dosud vydanou publikaci. Pro usnadnení a zprehlednení práce jsem si tedy stanovil další dílcí cíle: Prostrednictvím casopisu postihnout a rozkrýt události v CSM a studentském hnutí. Priblížit fungování cenzury a její vliv na novinárskou práci. Popsat nástup nové generace mladých novináru druhé poloviny šedesátých let a její konfrontaci se starší generací. Vystihnout vztah casopisu Student s Literárními novinami. Postihnout význam casopisu Student behem krize ceskoslovenské spolecnosti v roce 1967, v období "Pražského jara" a následné okupace vojsky Varšavské smlouvy. Popsat životní osudy nejvýznacnejších osob spojených s casopisem Student

Induction of MAPK signaling by SMOC does not require the EGF receptor.

<p>Immunoblot analysis of serum-starved HEK293 (A, D) or 32D/EGFR (B, C) cell lysates following addition of <i>X</i>SMOC-1 (100μg/ml) or hSMOC-1 (50μg/ml). (A) Following addition of <i>X</i>SMOC-1 to HEK293 cells, phosphorylation of MEK1/2 and Erk was evident at four minutes. (B) Phosphorylation of Erk and EGFR (Y1172) in 32D/EGFR cells was weak in the presence of <i>X</i>SMOC-1 compared to hSMOC-1. (C) Phosphorylation of Erk by hSMOC-1 in 32D/EGFR cells was blocked by the small molecule EGFR inhibitor, Gefitinib (5μM). (D) Phosphorylation of Erk by SMOC in HEK293 cells does not require the EGFR; <i>X</i>SMOC-1 and hSMOC-1 continue to induce Erk phosphorylation in the presence of Gefitinib but, in the absence of Gefitinib, hSMOC-1 stimulated ERK phosphorylation more than <i>X</i>SMOC-1. Non-phosphorylated Erk and EGFR are shown as loading controls. The results presented are representative of experiments conducted at least three times.</p

Pro-EGF is present as an impurity in a commercial hSMOC-1 product.

<p>(A) Immunoblot of 5μg hSMOC-1 (R&D systems #6074-SM) with an antibody to mature EGF (EMD Millipore #PC08). The band detected at approximately 160 kDa is consistent with the mass of pro-EGF. Mature EGF (50ng), detected at 5kDa, is shown as control. (B) Immuno-quantitation of pro-EGF in hSMOC-1. Fluorescence scan analysis of pro-EGF signals obtained following immunoblotting of hSMOC-1 (R&D Systems) and recombinant pro-EGF (R&D Systems #4289-EG) at the amounts shown. The left-hand Y axis displays the pro-EGF fluorescence signal intensity; the X axis displays the amounts of commercial hSMOC-1 analyzed (blue squares); the right hand Y axis displays the known amounts of recombinant pro-EGF analyzed (red squares). Note: Additional pro-EGF data points were used for the best fit analysis, but the graph was cropped to show the intersect point with hSMOC-1 more clearly. (C) Immunoblot of 32D/EGFR cell lysates showing increased phosphorylation of Erk following a six minute exposure to hSMOC-1 or hSMOC-1 immuno-depleted of pro-EGF (hSMOC-1 –proEGF). (D) Immunoblot of HEK293 cell lysates showing Erk phosphorylation following a six minute exposure to <i>X</i>SMOC-1, hSMOC-1, or hSMOC-1 –proEGF; the apparent higher potency of undepleted hSMOC-1 was presumably due to the pro-EGF impurity. The cell culture experiments were conducted in triplicate and the results presented are representative of those obtained.</p

Human pro-EGF peptide sequences obtained from MS-MS sequencing (ITSI Biosciences) of human SMOC-1 (R&D Systems).

<p>Human pro-EGF peptide sequences obtained from MS-MS sequencing (ITSI Biosciences) of human SMOC-1 (R&D Systems).</p

<i>X</i>SMOC-1 binds to pro-EGF and co-localizes with pro-EGF <i>in vivo</i>.

<p>(A) Immunoblot of pro-EGF following co-immunoprecipitation of pro-EGF with <i>X</i>SMOC-1, <i>X</i>SMOC-1 ΔEC, or <i>X</i>SMOC-1 EC in the presence of TBST/0.1% SDS; pro-EGF binds to <i>X</i>SMOC-1 and <i>X</i>SMOC-1 ΔEC, but not <i>X</i>SMOC-1EC. (B) RT-PCR analysis of HEK293 cells showing positive signal for pro-EGF(C) Representative confocal image showing co-localization of <i>X</i>SMOC-1 and pro-EGF (red fluorophore) on HEK293 cells using the PLA method. Nuclei are stained blue with DAPI. (D, E) Representative whole mount hybridization <i>in situ</i> images of <i>Xenopus</i> neurula embryos (stage 26) stained for <i>X</i>SMOC-1 (C) or pro-EGF (D). The locations of the eye (e) and pronephros (pn) are indicated.</p