DOI: 10.5061/dryad.j0zpc86sq
title: Supplementary data from: Critical role of LdZIP7 in enhancing cadmium tolerance of Lymantria dispar larvae: Functional identification at both the individual and cellular levels

description: Heavy metals are prevalent environmental pollutants in habitats of
 phytophagous insects. This study investigates the tolerance of Lymantria
 dispar larvae to cadmium (Cd) and the associated mechanisms
 involving the ZIP family. Cd stress reduced larval body weight and
 extended the development duration without causing significant mortality. A
 significantly up-regulated expression of apoptosis genes Caspase-1,
 Caspase-3, and Caspase-7 was observed in Cd-exposed larvae. In Cd-exposed
 larvae, expression of the apoptosis-inhibitory factor Bcl-2 in the
 mitochondrial pathway decreased, while apoptosis-inducing factors Bax and
 AIFM increased. Cd stress markedly elevated the expression of CHOP and
 Bip, key genes in the endoplasmic reticulum stress pathway. Among the ZIP
 family genes, LdZIP7 showed the highest up-regulation in response
 to Cd treatment. Silencing LdZIP7 intensified the negative
 impacts of Cd stress on L. dispar larvae and significantly
 reduced the tolerance of L. dispar larvae to Cd. The main
 manifestations were a further significant decrease in larval body weight,
 a further significant extension of developmental duration, and the further
 activation of the mitochondrial pathway and the endoplasmic reticulum
 stress pathway-triggered apoptosis in
 Cd-treated larvae. At the Sf9 cell level, LdZIP7
 predominantly localizes in the nuclear membrane and cell
 membrane. Overexpression of LdZIP7 mitigates Cd-induced
 cytotoxicity by inhibiting the Ca2+-MPTP opening degree-mitochondrial
 membrane potential-apoptosis pathway. Overall, LdZIP7 plays a
 pivotal role in alleviating the biotoxic effects of Cd and is a
 significant regulatory gene for Cd tolerance in L. dispar larvae.
description: # Critical role of LdZIP7 in enhancing cadmium tolerance of Lymantria
 dispar larvae: Functional identification at both the individual and
 cellular levels: Open Source Data --- We have submitted our raw data
 (**Data.xls**) and a folder of unmodified fluorescence micrograph images
 (**Figure**). ## Variable Descriptions ### Data.xls * *C*: control group *
 *LCd*: Low Cd concentration treatment group * *HCd*: High Cd concentration
 treatment group * *Cd+LdZIP7*: Sf9 cells transfected with the recombinant
 plasmid pAC5.1b-EGFP-2/LdZIP7 served as the experimental subjects *
 *Cd+EGFP*: Untreated Sf9 cells and those transfected with EGFP were
 utilized as negative controls * *Figure 1A*: The weight of the larvae at
 different Cd concentrations (g) * *Figure 1B*: The developmental duration
 of the larvae under different Cd concentrations (h) * *Figure 1C*: The
 mortality rate of the larvae under different Cd concentrations (%) *
 *Figure 2*: The expression level of the ZIP gene family genes in the
 midgut of larvae * *Figure S1*: The gene expression level after silencing
 LdZIP7 * *Figure 3A*: The weight of the 4th instar larvae in different
 treatment groups after silencing LdZIP7 (g) * *Figure 3B*: The weight of
 the 5th instar larvae in different treatment groups after silencing LdZIP7
 (g) * *Figure 3C*: The developmental duration of the larvae in different
 treatment groups after silencing LdZIP7 (h) * *Figure 3D*: The mortality
 rate of the larvae in different treatment groups after silencing LdZIP7
 (%) * *Figure 4*: Subcellular localization of LdZIP7 protein in Sf9 cells
 * *Figure S2*: Relative expression levels of growth regulatory genes in
 Cd-treated larvae after LdZIP7 silencing * *Figure S3*: Relative
 expression levels of apoptosis indicator genes in Cd-treated larvae after
 LdZIP7 silencing * *Figure S4*: Relative expression levels of
 mitochondrial apoptosis pathway genes in Cd-treated larvae after LdZIP7
 silencing * *Figure S5*: Relative expression levels of endoplasmic
 reticulum stress pathway genes in Cd-treated larvae after LdZIP7 silencing
 * *Figure S7*: The survival rate of Sf9 cells under Cd stress * *Figure
 5A*: Cell viability of different treatment groups (%) * *Figure 5B*: ROS
 level of different treatment groups (%) * *Figure 5C*: Fluorescence images
 of ROS in different groups * *Figure 6A*: Fluorescence images of Ca2+ in
 different groups * *Figure 6B*: Diagram of Sf9 cell apoptosis in different
 treatment groups * *Figure 6C*: Analysis of Ca2+ fluorescence intensity of
 different treatment groups (%) * *Figure 6D*: Apoptosis rate of different
 treatment groups (%) * *Figure 7A*: The red/green fluorescence
 representing the MMP * *Figure 7B*: The green fluorescence representing
 the MPTP * *Figure 7C*: Analysis of MMP of different treatment groups (%)
 * *Figure 7D*: Analysis of MPTP of different treatment groups (%) ##
 Code/Software Throughout the entire project, the following tasks were
 carried out using BioRad, ImageJ, Prism, and SPSS software, respectively:
 1) Data collection, 2) data analysis, 3) graph creation, and 4) providing
 annotations.