IDEAS Quiz 2

spatial calibration

used to estimate total # of cells in dish

confluence

what is the coverage of the surface area of the cells

monolayer

contact inhibition; will have single layer of cells that do not stack up anymore & become quiescent

quiescent

not proliferate anymore

phenotype

array of proteins expressed by cells; determines how cells behave/appear

cell characteristics

morphology, growth behavior

proteins

during growth, quiescense

genes

encode proteins, if you want more protein upregulate this; during growth > quiescence

function of cell growth

time, media, density, cell type, extra cellular matrix, stimulus

how do monolayers stay adhered to dish

focal contacts are formed by endothelial cells (or focal adhesions)

integrin proteins

proteins that adhere to extracellular matrix

modes of transduction

mechanical, electrical, biochemical

time scales

immediate, positive/negative feedback, gene regulation

types of signaling

ECM interactions & synthesis; autocrine and paracrine communication; cell-cell interactions and gap junction communication

MAPK

Mitogen activated protein kinase

MAPKs stimulate

cell growth, proliferation, survival, differentiation, migration

kinases

proteins that serve as an enzyme

phosphorylated form of kinases

phosphate is added to phosphate dependent section of protein where there is conformational change

protein phosphate

counteracts kinase

ERK

Extracellular Regulated Kinase

downstream gene

EGR-1

MAPKK

Mitogen-Activated Protein Kinase Kinase

confluence

degree (5) to which cells have covered the available area

as coverage by cells increases, what happens to growth rate?

it decreases

term for phenomenon where growth rate decreases as coverage increases

contact inhibition

In the RNA purification procedure, what is the final liquid that will be eluted through the column (and in which the RNA will be dissolved)

RNase-free water

Time for the signaling protein and gene probing

5 minutes to serum stimulus for ERK to reach peak phosphorylation

Time for the signaling protein and gene probing step 2

30 minutes so EGR-1 mRNA is high enough to detect & gene transcription occurs

how the silica-gel-based membranes in the Qiagen "RNeasy" kit selectively enrich for mRNA (as opposed to tRNA or rRNA).

only RNA above 200 nucleotides are isolated and stick to column but only mRNAs fit this length

What does a lysis buffer do?

it breaks down cells allowing them to release their contents