Journal: Molecular Systems Biology

Article Title: An immediate–late gene expression module decodes ERK signal duration

doi: 10.15252/msb.20177554

Figure Lengend Snippet: Upper panel: pERK2 log 2 fold changes upon different input scenarios (sustained: 4OHT; 2‐h pulse: 4OHT + U0126). Lower panel: response amplitude across temporal clusters and signal durations. Bold lines show median cluster amplitude at each time point. Capacity to decode signal duration. ILGs translate ERK signal duration into response amplitude. IEGs are only partially able to do so, as many of these genes are still strongly induced upon shortened signal durations. qPCR validation to test different ERK signal durations. HEK293∆RAF1:ER cells were treated with 4OHT and U0126 for different periods of time to generate signal duration scenarios of 0.5–8 h (cf. Fig C). mRNAs of IEGs EGR1 and FOS relay signal duration to response duration, whereas ILGs CLU and FOSL1 decode signal duration to response amplitude (qPCR data for all 17 validated mRNAs is shown in Fig A). Relation between signal duration and response amplitude for IEGs and ILGs derived from qPCR validation data. Median amplitude is based on six qPCR‐validated ILGs and six qPCR‐validated IEGs. Quantification of Western blots to present protein log 2 fold changes of sample genes upon sustained ERK signalling (4OHT‐induced) and transient ERK signalling (EGF‐induced) in HEK293∆RAF1:ER cells. Conservation of signal duration decoding to response amplitude in two prominent model systems for ERK signal duration: NGF and HRG cause more sustained ERK signalling compared to EGF treatment in PC12 and MCF7 cells, respectively. Decoding of signal duration to response amplitude is clearly governed by ILGs.

Article Snippet: Western blot antibodies were as follows: EGR1 (Santa Cruz sc‐110), FOS (Cell Signaling #2250), CLU (Santa Cruz sc‐8354), FOSL1 (Santa Cruz sc‐376148).

Techniques: Biomarker Discovery, Derivative Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Insulin Treatment Forces Arteriogenesis in Diabetes Mellitus by Upregulation of the Early Growth Response-1 (Egr-1) Pathway in Mice

doi: 10.3390/ijms20133320

Figure Lengend Snippet: Gene expression studies of ( a ) mRNA levels and ( b ) protein levels of collateral arteries obtained from occluded and sham-operated mice 24 h after the surgical procedure. ( a ) Dot plots representing the results of qRT-PCR analyses ( n = 5 per group, * p < 0.05 (each group compared to each other group), # p < 0.05 compared to corresponding occ group from one-way ANOVA with Bonferroni’s multiple comparison test). Results were normalized to the expression level of the 18S rRNA. ( b ) Quantitative analyses (upper panel) and corresponding representative pictures of a Western blot (lower panel) showing the protein expression of Egr-1 as well as of α-tubulin, which was used for normalization, 24 h post-FAL ( n = 4 per group, * p < 0.05 (each group compared to each other group) from one-way ANOVA with Bonferroni’s multiple comparison test).

Article Snippet: The blots were probed for Egr-1 protein (Egr-1 antibody (588)-Santa Cruz BioTechnology Cat.no #sc110). α-Tubulin served as a housekeeping protein (alpha tubulin antibody (B-7): Santa Cruz Biotechnology Cat.no.

Techniques: Expressing, Quantitative RT-PCR, Western Blot