RAW DATA README FOR Data Accessibility for Manuscript MEC-14-0904.R1 :
Spatial analysis of gene regulation reveals new insights into the molecular basis of upper
thermal limits.
Marina Telonis-Scott, Allannah S. Clemson, Travis K. Johnson and Carla M. Sgrò
DATAFILE: Raw_heat_knockdown_data.csv
Column A = Individual: individual fly
Column B = Block: Knockdown data was generated from three replicate blocks of approximately
35 flies totaling at least 100 individuals per treatment/population.
Column C = Thermal regime: ‘basal’= exposure to static heat stress at 38.5°C or ‘hardened’ =
pre-exposure to static heat 37°C for 1 hour with a 6-hour recovery on media, followed by
exposure to static severe stress at 38.5°C.
Column D = Population: D. melanogaster mass-bred populations derived from three locations
along the Australian East Coast representing high, mid, and low latitudes; Melbourne (37.99 oS,
145.27oE), Port Macquarie (30.93oS, 152.90oE), and Townsville (19.26oS, 146.79oE) respectively.
Column E = Latitude: Melbourne (37.99oS, 145.27oE), Port Macquarie (30.93oS, 152.90oE), and
Townsville (19.26oS, 146.79oE) respectively.
Column F = Heat Knockdown time in minutes (time to incapacitation).
Column G = log heat knockdown time = natural log of knockdown time.
DATAFILE: stv_rawcp_with_RPL11_unfiltered_data.csv
Note this is the complete unfiltered, un-normalized data set including data-points that were
later excluded in the analyses such as data points with technical replicates showing a standard
deviation >1, and the circadian controls (see methods).
Column A = Gene: either RPL11 (normalizer) or stv (gene of interest).
Column B = Transcript: either RPL11 or stv isoforms. Note, the corresponding RPL11 sample for
each transcript is denoted as RPL11_’transcript’.
Column C = Thermal_regime: ‘basal’= exposure to static heat stress at 38.5°C or ‘hardened’ =
pre-exposure to static heat 37°C for 1 hour with a 6-hour recovery on media, followed by
exposure to static severe stress at 38.5°C.
Column D = Population: D. melanogaster mass-bred populations derived from three locations
along the Australian East Coast representing high, mid, and low latitudes; Melbourne (37.99 oS,
145.27oE), Port Macquarie (30.93oS, 152.90oE), and Townsville (19.26oS, 146.79oE) respectively.
Column E = Time-point_hours= sampling time-points for the transcript expression time-course,
either 0 = time_zero, or pre-stress, 0.15h = 15 minutes of heat shock, 0.53 = 31.5 minutes of
heat shock, 4h= 4 hours recovery post-heat shock, 8h= 8 hours recovery post-heat shock, 12h=
12 hours recovery post-heat shock, 24h= 24 hours recovery post-heat shock, 48h= 48 hours
recovery post-heat shock, or 12, 24 48h = circadian controls (received no heat shock).
Column F = Temperature_degreesC = temperature flies were sampled at, either 38.5°C or 25°C.
Column G = Treatment = denotes the treatment for each sample, either basal or hardened,
heat shock or recovery, or untreated flies sampled at 12, 24, and 48h post stress as circadian
rhythm controls (see methods).
Column H = Replicate, 1, 2 and 3 denotes biological replicate. In this file there are also two
technical replicates for each sample to control for measurement error on the Lightcycler. Each
biological replicate appears twice.
Column I = raw_cp: raw transcript expression as determined by the Roche Lightcycler 480
software.
DATAFILE:stv_delatcp_relative_to_RPL11.csv
Note this is normalized data set where delta_cp is the relative expression of the stv isoforms to
the reference or ‘housekeeper’ RPL11. First technical replicates where averaged, and filtered
for those measures which had a standard deviation >1, then where relative expression of
transcript of interest (TOI) = 2^(Rpl11-TOI).
Column A = Transcript: stv-isoforms
Column B= Thermal_regime: ‘basal’= exposure to static heat stress at 38.5°C or ‘hardened’ =
pre-exposure to static heat 37°C for 1 hour with a 6-hour recovery on media, followed by
exposure to static severe stress at 38.5°C.
Column C = Population: D. melanogaster mass-bred populations derived from three locations
along the Australian East Coast representing high, mid, and low latitudes; Melbourne (37.99 oS,
145.27oE), Port Macquarie (30.93oS, 152.90oE), and Townsville (19.26oS, 146.79oE) respectively.
Column D = Time-point_hours= sampling time-points for the transcript expression time-course,
either 0 = time_zero, or pre-stress, 0.15h = 15 minutes of heat shock, 0.53 = 31.5 minutes of
heat shock, 4h= 4 hours recovery post-heat shock, 8h= 8 hours recovery post-heat shock, 12h=
12 hours recovery post-heat shock, 24h= 24 hours recovery post-heat shock, 48h= 48 hours
recovery post-heat shock, or 12, 24 48h = circadian controls (received no heat shock).
Column E = Temperature_degreesC = temperature flies were sampled at, either 38.5°C or 25°C.
Column F = Treatment = denotes the treatment for each sample, either basal or hardened,
heat shock or recovery, or untreated flies sampled at 12, 24, and 48h post stress as circadian
rhythm controls (see methods).
Column G = Replicate, 1, 2 and 3 denotes biological replicate. Note this is the average of the
two technical replicates.
Column H = delta_cp_relative _to_RPL11: relative expression of transcript of interest to RPL11.