B6n 129s prom1tm1 cre ert2 gilb j

Hepatocyte-specific Shp2 knockout (Alb-Cre; Shp2^flox/flox) or SKO mice were generated as described previously (Bard-Chapeau et al., 2011 (link)). c-Met^flox/flox mice (FVB;129P2-Met^tm1Sst/J) were purchased from the Jackson Laboratory and bred with Alb-Cre mice to produce hepatocyte-specific c-Met KO mice. Prom1 KO mice (B6N;129S-Prom1^{tm1(cre/ERT2)Gilb}/J) were purchased from the Jackson Laboratory and bread with Shp2^flox/flox and SKO mice. Shp2^flox/flox littermates were used as WT control mice. All experimental procedures were approved by the UCSD Institutional Animal Care and Use Committee (IACUC). PHx was performed on mice at age of 8–10 weeks with the use of Buprenorphine hydrochloride (Par Pharmaceutical) as a pain relief for the surgery. CCl₄ (20% in 100 μl corn oil/20 g of body weight; Sigma) was intraperitoneally injected into 8–10 week-old mice. AAV-Cre (AAV8.TBG.PI.Cre.rBG) was purchased from the Penn Vector Core and injected into the tail vein (2x10¹¹/28 gbw). For chemical induction of hepatocellular carcinoma (HCC), DEN (25 mg/kg; N0258-1G; Sigma) was injected intraperitoneally into mice at postnatal day 15. BrdU (1 mg/mice; Sigma) was intraperitoneally injected twice a day for 3 days.

Hepatocyte-specific Shp2 knockout (Alb-Cre; Shp2^flox/flox) or SKO mice were generated as described previously (Bard-Chapeau et al., 2011 (link)). c-Met^flox/flox mice (FVB;129P2-Met^tm1Sst/J) were purchased from the Jackson Laboratory and bred with Alb-Cre mice to produce hepatocyte-specific c-Met KO mice. Prom1 KO mice (B6N;129S-Prom1^{tm1(cre/ERT2)Gilb}/J) were purchased from the Jackson Laboratory and bread with Shp2^flox/flox and SKO mice. Shp2^flox/flox littermates were used as WT control mice. All experimental procedures were approved by the UCSD Institutional Animal Care and Use Committee (IACUC). PHx was performed on mice at age of 8–10 weeks with the use of Buprenorphine hydrochloride (Par Pharmaceutical) as a pain relief for the surgery. CCl₄ (20% in 100 μl corn oil/20 g of body weight; Sigma) was intraperitoneally injected into 8–10 week-old mice. AAV-Cre (AAV8.TBG.PI.Cre.rBG) was purchased from the Penn Vector Core and injected into the tail vein (2x10¹¹/28 gbw). For chemical induction of hepatocellular carcinoma (HCC), DEN (25 mg/kg; N0258-1G; Sigma) was injected intraperitoneally into mice at postnatal day 15. BrdU (1 mg/mice; Sigma) was intraperitoneally injected twice a day for 3 days.

The Prom1 rd19 mice were acquired from the Jackson laboratory (B6. BXD83-Prom1rd19/Boc, RRID:IMSR_JAX:026803). These rd19 animals are in C57Black/6J background and carry a hypomorphic RPE65 mutation with methionine at position 450 (Rd19 M/M). To generate a mouse line with the RPE65 allele coding for leucine at position 450, we crossed the rd19 mice with 129SVE/J mice. This line, referred to as PROM1 knockout, is used throughout this study unless specified otherwise and is maintained in a mixed background. The mice were housed in the West Virginia University animal facility with a 12-h light/dark cycle with ad libitum access to food and water.
For dark rearing, mating pairs were raised in complete darkness, without any light. In dark rearing rooms, mice were placed in a temperature- and humidity-controlled incubator (Powers Scientific Inc., cat# HIS33SD). The glass doors of the incubators were covered with light-insulating material to ensure complete darkness. Temperature and humidity were monitored daily by the lab personnel, and red light was used for maintenance tasks. Prom1 Cre-ERT2 mice (B6N;129S-Prom1tm1(cre/ERT2)Gilb/J, RRID:IMSR_JAX:017743) and rd10 mice (B6.CXB1-Pde6brd10/J, RRID:IMSR_JAX:004297) were obtained from Jackson Laboratories.