DOI: 10.5061/dryad.nzs7h451x
title: Raw Western blots data to assess cold shock protein expression

description: Preclinical and clinical studies show that mild to moderate hypothermia is
 neuroprotective in sudden cardiac arrest, ischemic stroke, perinatal
 hypoxia/ischemia, traumatic brain injury and seizures. Induction of
 hypothermia largely involves physical cooling therapies, which induce
 several clinical complications, while some molecules have shown to be
 efficient in pharmacologically-induced hypothermia (PIH). Neurotensin
 (NT), a 13 amino-acid neuropeptide that regulates body temperature,
 interacts with various receptors to mediate its peripheral and central
 effects. NT induces PIH when administered intracerebrally. However, these
 effects are not observed if NT is administered peripherally, due to its
 rapid degradation and poor passage of the blood brain barrier (BBB). We
 conjugated NT to peptides that bind the low-density lipoprotein receptor
 (LDLR) to generate “vectorized” forms of NT with enhanced BBB
 permeability. We evaluated their effects in epileptic conditions following
 peripheral administration. One of these conjugates, VH-N412, displayed
 improved stability, binding potential to both the LDLR and NTSR-1,
 rodent/human cross-reactivity and improved brain distribution. In a mouse
 model of kainate (KA)-induced status epilepticus (SE), VH-N412 elicited
 rapid hypothermia associated with anticonvulsant effects, potent
 neuroprotection and reduced hippocampal inflammation. VH-N412 also reduced
 sprouting of the dentate gyrus mossy fibers and preserved learning and
 memory skills in the treated mice. In cultured hippocampal neurons,
 VH-N412 displayed temperature-independent neuroprotective properties. To
 the best of our knowledge, this is the first report describing the
 successful treatment of SE with PIH. In all, our results show that
 vectorized NT may elicit different neuroprotection mechanisms mediated
 either by hypothermia and/or by intrinsic neuroprotective properties.
description: Western blots were obtained using antibodies that detect RBM3 and
 CIRBP cold shock proteins and beta-actin used as an internal control, on
 hippocampal samples.
description: # Western blots
 [https://doi.org/10.5061/dryad.nzs7h451x](https://doi.org/10.5061/dryad.nzs7h451x) ## Description of the data and file structure Brains were extracted, cut in two for mRNA and protein analysis. In each hemi-section, the hippocampus was isolated and snap frozen. RT-qPCR analysis normalized with GAPDH on pooled samples showed significant increase of mRNA encoding RBM3 and CIRBP relative to PBS injected control at the 16 H and 8 H time points, respectively. In order to assess whether increased mRNA levels translate into increased protein levels in hippocampal samples, we used western blot and antibodies that detect RBM3, CIRBP and beta-actin used as an internal control. The raw data for the western blots is provided. ### Files and variables #### File: image2CIRBP\_PBS\_VH-N412\_4H\_8H\_16H\_Ladder.tif **Description:** Western blot to assess ladder size and the levels of CIRBP after VH-412 treatment at 4, 8, 16 hours.  #### File: subImage1\_b-actin\_PBS\_VH-N412\_4H\_8H\_16H.tif **Description:** Western blot to assess the levels of beta-actin used as an internal control, after VH-412 treatment at 4, 8, 16 hours. #### File: subImage11RBM3VH-N412\_PBS\_4H\_8H.tif **Description:** Western blot to assess the levels of RBM3 after VH-412 treatment at 4 and 8 hours. #### File: subImage9RBM3VH-N412\_16H.tif **Description:** Western blot to assess the levels of RBM3 after VH-412 treatment at 16 hours. #### File: subImage7CIRBP\_PBS\_VH-N412\_4H\_8H\_16H.tif **Description:** Western blot to assess the levels of CIRBP after VH-412 treatment at 4, 8, 16 hours. ## Code/software Image TIF, images can be visualized with Photoshop or Aperçu