Crthuky

I need to have a full-width image at the bottom of the page and a small, half-size image at the top. There will be text in between. I've tried messing around with the h,t,b,! of both images to no avail.

Here is my code:

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beginfigure[!t]
centering
includegraphics[width=1linewidth, height=5 cm]Figures/iVSMCstaining.png
captiontextbfStaining confirming iVSMC generation. VSMCs that were stained for smooth muscle aortic alpha-actin 2 (ACTA2) and 4′,6-diamidino-2-phenylindole (DAPI). Red corresponds to ACTA2 conjugation, and light blue corresponds to DAPI conjugation. Alexa-fluorophore 555 was the secondary antibody used to visualize the proteins of interest. Scale bar = 400 um.
labelfig:DriftCorrection
endfigure
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Given the qPCR analysis and staining results, it is apparent that our differentiation protocol resulted in the generation of iPSC-derived VSMCs.

subsection*E-protein constructs
Two methods were used to design the final E-protein CRISPR delivery systems; the formation of in-del mutations via non-homolgous end joining (NHEJ) CRISPR editing and base editor CRISPR editing. Each method used a multi-vector approach, with both methods making use of a pSaGuide Cas9 plasmid that contained our guide sequence that helped target the CRISPR constructs to our gene of interest. The NHEJ method uses the guide plasmid in combination with a plasmid that expresses a catalytically active version of cas9. The base editor method combines the guide plasmid with a second plasmid that fuses a catalytically dead Cas9 to a cytidine deaminase. A multi-vector approach was used in order to avoid having plasmids that were too large, and therefore unable to enter the iPSCs.

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beginfigure*[!h]
centering
includegraphics[width=textwidth]Figures/guideplasmiddesign.png
captiontextbfGuide Plasmid Design textbfA) The pSaGuide backbone was ordered from Addgene.The pSaGuide plasmid contains an ampicillin resistance (AmpR) cassette, a kanamycin/neomycin resistance (KanR/NeoR) cassette, a U6 promoter region, and the Sa gRNA scaffold. textbfB) The chart in the top corner represents the guide RNA oligonucleotide (oligos) sequences that were designed and ordered as well. The oligos were annealed together to form double stranded DNA (dsDNA) and phosphorylated to convert the deoxyribonucleotide diphosphates (DNDPs) to deoxyribonucleotide triphosphates (DNTPs). The conversion from DNDP's to DNTP's allows the DNA to get ligated into the cut pSaGuide backbone via the use of overhanging sticky ends. Each oligo has a sequence that is analogous to the ends of the cut plasmid, which allows for easy ligation and incorporation.
labelfig:GeneralDiagram
endfigure*
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